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PHYSIOLOGY 30: 438 – 448, 2015; doi:10.1152/physiol.00019.2015
Plasma Membrane Repair: A Central
Process for Maintaining Cellular
Homeostasis
Plasma membrane repair is a conserved cellular response mediating active
Alisa D. Blazek,* Brian J. Paleo,* and
Noah Weisleder
Department of Physiology and Cell Biology, Davis Heart and
Lung Research Institute, The Ohio State University Wexner
Medical Center, Columbus, Ohio
[email protected]
* A. D. Blazek and B. J. Paleo contributed equally to this review.
resealing of membrane disruptions to maintain homeostasis and prevent cell
death and progression of multiple diseases. Cell membrane repair repurposes
mechanisms from various cellular functions, including vesicle trafficking, exocytosis, and endocytosis, to mend the broken membrane. Recent studies
increased our understanding of membrane repair by establishing the molecthe key proteins linked to cell membrane repair.
A eukaryotic cell is separated from the extracellular environment by a plasma membrane composed of a phospholipid bilayer containing
proteins that regulate transit of molecules into
and out of the cell. Loss of this barrier function
can lead to compromised cellular homeostasis
and death of the cell. Most cells are subjected to
mechanical or chemical stresses that can disrupt
the plasma membrane; thus there is strong selective pressure to ensure the integrity of this
membrane. The inherent nature of phospholipids and early work with lysosomes indicated that
the plasma membrane would thermodynamically reseal after disruption (112). While this is
true of simple lipid bilayers or small membrane
disruptions (FIGURE 1A), the plasma membrane
contains integral proteins that interact with the
cytoskeleton and extracellular matrix to support
numerous cellular functions. These interactions
create mechanical tension on the plasma membrane that holds the membrane open after disruption (25, 114). Such disruptions allow intracellular
components to escape the cell and potentially permit toxic levels of Ca2⫹, oxidants, and other components of the extracellular milieu to enter the cell.
Thus, if these disruptions are not closed rapidly, it
may lead to the death of the cell. As a result, cells
have evolved active methods to reseal plasma
membrane disruptions in which normal cellular
responses are repurposed to mend the broken
membrane (70, 106, 130) through a process called
membrane repair.
The idea of facilitated membrane repair was supported by earlier work (23, 59) before the concept
was formally presented by McNeil and colleagues,
who initially showed that plasma membrane disruptions and repair occur in vivo and that damaged cells reseal by recruitment of intracellular
vesicles to form a repair patch in an extracellular
438
Ca2⫹-dependent manner (FIGURE 1B) (12, 96 –98,
106, 126). Membrane repair can also involve fusion
of vesicles at the injury site or into the proximal
plasma membrane. Constriction of the membrane
around disruptions can also contribute to membrane
repair (FIGURE 1C) (9). Endocytotic mechanisms
may be involved in resealing of larger membrane
disruptions, whereas smaller disruptions of ⬍100
nm reseal through budding and exocytosis
(FIGURE 1D). Repair through budding involves
pinching the membrane at the injured site and
shedding the injured membrane into the extracellular space (FIGURE 1E) (4, 74, 79). Endocytosis
is also thought to contribute to membrane repair
by internalization of the injured membrane
(FIGURE 1F) (70, 74). How and under what specific
conditions these mechanisms contribute to membrane repair is still an area of investigation. These
non-exclusive mechanisms could be relevant in a
cell-type and context-dependent fashion. It is clear
that compromised membrane repair contributes to
pathophysiology in a number of different tissues
and that it is linked to muscular dystrophy, heart
failure, neurodegeneration, and other diseases (5,
7, 24, 27, 50, 72, 133, 140). Despite the importance
of membrane repair in cellular function, the field
has only recently begun to expand with the discovery of more proteins linked to resealing damaged
cell membranes. The focus of this review is to
identify several proteins currently linked to membrane repair and to describe some of the key findings on their functions.
Ferlin Family (Dysferlin/Myoferlin/
Otoferlin)
Dysferlin is a type II transmembrane protein from
the ferlin family that localizes to the plasma membrane and T-tubules of muscle fibers (FIGURE 2)
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ular machinery contributing to membrane resealing. Here, we review some of
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Calpain
After membrane disruption, rapid reorganization
of cytoskeletal elements at the site of injury is
necessary. Treatment with the actin-depolymerizing enzyme DNase I significantly enhanced resealing, demonstrating that disassembly of the actin
skeleton is important in membrane resealing (107).
Calpain 3 is a cysteine protease that cleaves cytoskeletal proteins, such as talin and vimentin, and
may aid in early remodeling of various proteins
during membrane repair as cleaved fragments of
these cytoskeletal proteins could not be recovered
in calpain-null cells (101). Calpain 3 cleaves
AHNAK, inhibits AHNAK’s interactions with dysferlin and myoferlin, and regulates AHNAK protein
turnover (FIGURE 2). Through these actions, calpain regulates cytoskeletal structure and interaction of the cytoskeleton with the cell membrane
(34, 67). In fact, loss of calpain 3 in particular has
been shown to cause limb girdle muscular dystrophy (119). Calpain has been shown to be required
for calcium-dependent membrane repair, since
knockout of the small subunit of the calpain enzyme results in failure to reseal after laser-induced
membrane disruption (101). Calpain could contribute to membrane repair through other mechanisms as well. Studies suggest that calpain can
remodel sarcomeres since calpain 3 binds titin and
is present in other regions of the sarcomere, such
as the Z disk, costameres, and myotendinous
junctions (45). Thus the role of calpain may be to
increase local loosening of the sarcomere via proteolysis and facilitate removal of damaged and
cleaved proteins by the proteosome (8). Other
studies suggest a role for calpain 3 in post-membrane repair sarcolemmal remodeling because loss
of calpain in limb girdle muscular dystrophy leads
to disorganization of myofibers and a lack of organized sarcomeres and sarcomere proteins in myotubes (45, 67, 100). Finally, calpain activity leads to
dysferlin cleavage to produce subunits that function in membrane repair (116).
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(1, 2). Dysferlin was initially identified as the target
gene mutated in Myoshi Myopathy (MM) and
limb-girdle muscular dystrophy (LGMD) type 2B
(6, 71, 87, 132). Subsequent dysferlin knockout
mouse studies showed that muscle fibers (63) and
hearts (24, 56) from these animals demonstrated a
decreased ability to reseal the plasma membrane
(5), leading to muscular dystrophy and late-onset
cardiomyopathy (5, 24). These results identified
dysferlin as the first protein to contribute directly
to membrane repair in striated muscles, resulting
in dysferlin’s prominent role in the membrane
repair literature (26). Subsequent studies linked
dysferlin to roles in membrane receptor recycling,
endocytosis, vesicle trafficking, membrane turnover, focal adhesion, and ATP-dependent intercellular signaling and modulation of the immune
system (31, 34, 36, 47, 58, 109, 139, 141). Recent in
vivo studies indicated that dysferlin may function
in the maintenance of T-tubule structure (78) and
that the principal function of dysferlin may be at
the T-tubules rather than in sarcolemmal membrane (77). Additional studies will be necessary to
establish the extent that these findings alter the
current understanding of dysferlin in membrane
repair; however, it is clear that much is still unknown about dysferlin function.
Other members of the ferlin family also have
been linked to membrane repair. For example,
myoferlin shares a 56% homology to dysferlin and
is expressed during muscle development to facilitate myoblast fusion (41, 139). Wild-type mice show
low expression of myoferlin; however, increased expression was observed in mdx and gamma-sarcoglycan-null mouse models of muscular dystrophy (37,
141). Transgenic mouse experiments showed that
myoferlin overexpression could compensate for dysferlin in membrane repair; however, myoferlin does
not prevent all dystrophy symptoms (90). Another
dysferlin family member, otoferlin, regulates synaptic vesicle exocytosis in cochlear hair cells and may
be involved in membrane repair specifically within
these cells (75, 121).
MG53/TRIM72
Mitsugumin 53 (MG53/TRIM72) has been shown
to be a vital component of the membrane repair
machinery in several cell types (18, 21, 44, 80). It is
a member of the tripartite motif family of E3 ubiquitin ligases (TRIM72) that was originally identified
in an immunoproteomic library screen for muscle
enriched proteins (136). Although native MG53/
TRIM72 protein was initially thought to be found
only in skeletal and cardiac muscle (18), recent
studies have shown expression and membrane repair function in other tissues (44, 73, 80). Along
with native protein repair capacity, overexpression
of MG53/TRIM72 in cells that do not express
MG53/TRIM72 also shows protective effects
against membrane injury (137). MG53/TRIM72null mice display defective membrane repair (18),
progressive myopathy (18), and increased susceptibility to injury in the heart (21, 135), lungs (73,
80), and kidneys (44).
MG53/TRIM72 interacts with phosphatidylserine to associate with intracellular vesicles and
the inner leaflet of the plasma membrane (18)
(FIGURE 3). Once a cell is injured, MG53/TRIM72 is
thought to react to the oxidized extracellular environment entering the cell by forming higher molecular weight units (69). MG53/TRIM72-tethered
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Caveolin
Caveolin family (Cav-1, -2, and -3) proteins are
21- to 24-kDa integral membrane proteins enriched in invaginations of the plasma membrane
(caveolae) and are involved in membrane transport (82) (FIGURES 1C AND 3). Cav-3 is muscle
specific and has been most closely linked with
membrane repair. Mutations in Cav-3 cause
autosomal-dominant LGMD1C and autosomaldominant rippling muscle disease (AD-RMD)
(110). Cav-3 has been shown by co-immunoprecipitation to interact with MG53/TRIM72 and
dysferlin (20), and Cav-3 overexpression regulates
membrane fusion events by downregulating
MG53/TRIM72-induced membrane budding and
preventing development of filopodia-like structures
(19). Disruption of the Cav-3/dysferlin/MG53 complex can affect the localization and membrane repair
function of the other components. For example,
dominant negative Cav-3 mutations associated with
the development of muscular dystrophy have been
shown to cause retention of MG53/TRIM72 (20) or
dysferlin (61) in the Golgi apparatus and loss of
membrane repair capacity. However, other studies
using ultrastructural analysis of dysferlin trafficking
showed that dysferlin can still reach the plasma
membrane in the absence of Cav-3 but that it is
rapidly endocytosed (60).
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vesicles traffic to the membrane disruption, allowing vesicles to fuse and patch the injured membrane (18). This process appears to involve a
dysferlin and Caveolin-3 (Cav-3) containing complex that regulates repair in the sarcolemma (20,
134). A study using ballistic injury and super-resolution microscopy of human myotubes showed
that MG53/TRIM72 is recruited quickly to the
membrane (2 s), followed by dysferlin ⬃10 s postinjury (84). MG53/TRIM72 and dysferlin form a
lattice that fills the wound area with vesicles and
closes the wound (84).
Interestingly, when myotubes are injured in media containing exogenous recombinant human
MG53/TRIM72 protein (rhMG53), the protein can
be seen to localize to the injury site (137). This
mechanism is similar to intracellular MG53/
TRIM72 binding to exposed phosphatidylserine
(18) and does not require endogenous MG53/
TRIM72 or dysferlin (137). This association with
injury sites can increase membrane resealing (137).
Application of rhMG53 to animal models of various
diseases, such as muscular dystrophy (137), myocardial infarct (88), lung injury (73, 80), compartment syndrome (28), and acute kidney injury (44)
can reduce the pathology in these models. While
this property of the protein is dependent on the
phosphatidylserine binding capacity of rhMG53, it
is possible that other aspects of the protein function, such as regulation of intracellular signaling,
could also contribute to this ability to increase
membrane repair capacity (55, 76, 83, 88). Further
studies should determine the rhMG53 extracellular
mechanism of action and assess its potential as a
therapeutic agent.
Polymerase-1 and Transcriptase
Release Factor (PTRF/Cavin-1/
Cav-p60)
PTRF may aid in the formation of caveolae at the
plasma membrane since PTRF localizes to caveolae, expression of PTRF is sufficient for caveolae
formation, and loss of PTRF results in a reduced
number of caveolae and a dystrophic phenotype
(17). Experiments using fluorescence lifetime imaging showed that cholesterol is required for the
interaction between PTRF and caveolin. Since cholesterol depletion decreases the PTRF-caveolin interaction, this suggests that PTRF may be involved
in stabilizing the membrane curvature of caveolae
(62). Additionally, it was shown by immunoprecipitation that PTRF binds dysferlin and, in fact,
may be required for the correct localization of
dysferlin, since PTRF mutation results in decreased
dysferlin at the cell membrane (17). Knockdown of
PTRF by shRNA results in decreased membrane
repair capacity, whereas overexpression of PTRF
can rescue dystrophic muscle membrane repair
(143). During the membrane repair process, PTRF
binds to dysferlin and may anchor MG53/TRIM72
to cholesterol, since MG53/TRIM72 cannot bind
cholesterol unless PTRF is present (92, 143). In this
model, PTRF anchors MG53/TRIM72 by binding to
FIGURE 1. Models of the plasma membrane repair process
A: thermodynamic resealing occurs spontaneously due to tension produced by the disordered arrangement of the
membrane phospholipids at the open edge of the break. This process is the most likely route of resealing for membrane breaks of ⱕ1 ␮m in diameter. B: exocytosis can contribute by trafficking intracellular vesicles to the wounded
area where they can fuse with each other and the injured membrane to form a repair patch. C: wound constriction is
mediated by caveolae. During this process, caveolae cluster and fuse around larger wounds, leading to wound constriction and intracellular fusion of caveolar endosomes. D: budding/blebbing of the membrane portion containing
the wound site with release of the newly formed vesicles into the extracellular space also involves exocytosis. E: exocytosis of an intracellular patch and fusion to the wound site could result in the extracellular release or “shedding” of
the wound site. F: endocytosis of wounds occurs via invagination of caveolar vesicles and subsequent intracellular fusion of caveolae.
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Affixin [␤-Parvin/Integrin-Linked
Kinase (ILK)-Binding Protein]
Motor Proteins (Nonmuscle Myosin
IIA and IIB/Kinesin)
Affixin was first discovered to be an integrinlinked kinase binding protein that localizes to
focal adhesions and likely contributes to their
maturation. In muscle cells, affixin and ILK colocalize at sites where the Z band attaches to the
sarcolemma due to an interaction with dysferlin
(94). Affixin’s interaction with dysferlin and altered immunoreactivity in MM, LGMD2B, and
LGMD1C suggest a role for this protein in muscle
membrane repair, possibly through organization
of F-actin (FIGURE 2A) (94). For example, affixin
has been shown by immunoprecipitation and
pull-down assay to interact with guanine nucleotide exchange factor ␣PIX (ARHGEF6 or Cool2), which functions to regulate actin skeleton
adhesion (93, 94, 104) as well as ␣-actinin, which
plays a role in organization of the cytoskeleton
(142). Although it appears that affixin may participate in cytoskeletal remodeling during mem-
Since vesicle trafficking is an important aspect of
membrane repair, motor proteins must be involved in this process to allow trafficking to occur.
Membrane repair is sensitive to inhibitors of both
myosin and kinesin motor proteins (126, 131).
Studies have specifically shown that non-muscle
myosin IIA and B are important motor proteins
that mediate vesicle trafficking during membrane
repair (131). Antisense knockdown of myosin IIB
suppressed exocytosis and membrane resealing,
and knockdown of myosin IIA inhibited the rate of
resealing at repeated wound sites. These studies
are supported by the observation that non-muscle
myosin IIA facilitates the transport of vesicles containing MG53/TRIM72 to the site of membrane
injury (86) (FIGURE 3).
FIGURE 2. Major membrane repair proteins and their hypothesized roles in the repair process
Dysferlin’s interaction with AHNAK is regulated by calpain, and cleavage of dysferlin can result in additional subunits that function in repair. Calpain may also regulate cytoskeletal structure and sarcomere remodeling. AHNAK
may aid in cytoskeletal remodeling. MG53/TRIM72 and dysferlin form a vesicle lattice to close the wound. A: affixin, which also binds dysferlin, localizes to focal adhesions and may organize actin. B: ESCRT and acid sphingomyelinase (ASM) facilitate exocytosis and endocytosis. ESCRT has been found to be involved in both endocytosis and
budding. ESCRT III can be recruited to the membrane, followed by blebbing of the membrane and shedding of
the wound. ASM is secreted and cleaves sphingomyelin to generate ceramide, leading to membrane invagination
of the injury site. C: the annexin and S100A10 complex binds dysferlin and may recruit AHNAK to the membrane
due to annexin’s ability to bind lipid rafts. Annexin/S100A10 may also bridge adjacent phospholipids to form endosomes. Annexin accumulates at the neck of membrane blebs to mediate microvesicle release. Annexin A6 may also
“cap” the membrane repair patch. D: synaptotagmin and SNARE proteins interact at the plasma membrane via a
conformational change in synaptotagmin present on synaptic vesicles to fuse the vesicles with the membrane.
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exposed cholesterol at the membrane injury site
(FIGURE 3).
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brane repair, the precise role of affixin in this
process remains speculative.
Acid Sphingomyelinase
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Acid sphingomyelinase (ASM) is an enzyme that
cleaves the phosphorylcholine head of sphingomyelin to generate ceramide, a molecule that leads to
membrane invagination, and that contributes to
the process of endocytosis during membrane repair (64, 124). In response to injury, lysosomes fuse
with the injured cell membrane and release ASM
from the cell. The action of ASM causes invagination of the membrane and endocytosis of the injury site (FIGURE 2B) (29, 35, 42, 129). Additional
evidence of ASM contribution to membrane repair
is provided by the finding that dysferlin-deficient
C2C12 cells showed less secretion of ASM than
control cells (35). When ASM was inhibited and
cells were permeabilized using a pore-forming
toxin, the cells could not sufficiently repair (35).
Treatment with exogenous ASM was sufficient to
restore the membrane integrity of ASM-depleted
cells (35) and of a dystrophic patient-derived myoblast cell line (35).
bind lipids while in their Ca2⫹-bound conformation, and the concentration of the Ca2⫹ signal determines which particular annexin family member
will bind to the phospholipid (53). Although the
exact mechanism remains unclear, numerous
studies suggest roles for annexins in vesicle movement, fusion, and patch formation during membrane repair (54, 57, 95). Annexins A1 and A2 bind
dysferlin in a Ca2⫹-dependent manner and may
contribute to membrane repair by their ability to
assist in the aggregation and fusion of intracellular
vesicles by their association with lipid rafts of the
plasma membrane (3, 85). Density gradient centrifugation experiments confirmed the Ca2⫹- and
annexin-dependent association of these rafts. Electron microscopic evidence shows that annexin
may be involved in membrane fusion during exocytosis as well as serving as a scaffold for endosomes (FIGURE 2C) (11, 53). Annexins also have
been shown to mediate microvesicle release and
blebbing, an indirect repair mechanism that involves sealing off a damaged membrane segment
by accumulation of annexin at the neck of the
membrane bleb (43). Other studies indicate that
annexin A5 (AnxA5) can bind to injured membranes and form a two-dimensional array that is
ESCRT
Endosomal sorting complex required for transport
(ESCRT) is involved in viral budding, cytokinesis,
and spontaneous budding of the plasma membrane. ESCRT subunits are classified into five complexes (108). The ESCRT III complex recently has
been shown to be involved in endocytosis and
budding in response to membrane damage (30, 74,
122). After injury, apoptosis-linked gene (ALG)-2
binds Ca2⫹ and leads to the accumulation of ESCRT III and accessory proteins ALG-2-interacting
protein X (ALIX), and vacuolar protein sorting-associated protein 4 (Vps4) (122). Assembly of this
complex results in cleavage of the wounded membrane and shedding of the wound site into extracellular vesicles. In response to UV laser injury,
ESCRT III is recruited to injury sites exercising
calcium-dependent wound repair. The observed
recruitment of ESCRT is followed by blebbing of
the membrane and shedding of the wound
(FIGURE 2B). ESCRT III is involved in endocytosis
of the membrane after the insertion of the bacterial
pore-forming toxin SLO (74). The complex appears
to be critical for repair of injuries of ⬍100 nm (74).
Annexin
Annexins are a protein family consisting of a
COOH terminus containing phospholipid and
Ca2⫹ binding sites, and a variable NH2 terminus
(111). The annexins are unique in that they can
myosin
FIGURE 3. Proposed roles of MG53/TRIM72 in mediating
membrane repair
MG53/TRIM72 interacts with phosphatidylserine in the plasma
membrane in a complex containing dysferlin and Cav-3. MG53/
TRIM72 and dysferlin close the membrane wound with vesicles. Vesicle transport is facilitated by myosin motor proteins. Cav-3 may
regulate MG53/TRIM72-mediated membrane fusion and is enriched
in caveolae or plasma membrane invaginations. PTRF may aid in the
formation and stabilization of these caveolae by interaction with
Cav-3 and MG53/TRIM72 through cholesterol. PTRF also binds and
may help localize dysferlin.
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thought to contribute to membrane repair, since
AnxA5-null cells show defective repair capacity
(15). Finally, annexin A6 has been shown by live
imaging to locate to membrane disruptions and to
assemble into a “cap” on the membrane repair
patch to assist with membrane resealing (128).
AHNAK (Desmoyokin)
S100A10
S100A10 is a small, ⬃10-kDa EF-hand Ca2⫹ binding protein also known as annexin 2 light chain or
p11. S100A10 forms a heterotetrameric complex
with annexin A2 by forming an S100A10 dimer in
the middle of two annexin A2 chains. This complex
targets to the plasma membrane in a calciumregulated manner that is dependent on its interaction with annexin 2 (FIGURE 2C). At the plasma
membrane, S100A10 interacts with a number of
proteins, where it may be essential for surface presentation of proteins such as ion channels (117).
Furthermore, proteomic and structural analyses
have identified that the heterotetrameric complex
binds cytosolic proteins AHNAK and dysferlin, and
that it is responsible for the recruitment of AHNAK
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Synaptotagmin/SNAREs
Synaptotagmins (Syt) are a family of proteins that
contain two C2 domains (C2A and C2B), which
some members use to bind Ca2⫹ or phospholipids
(16, 65, 102, 123, 127). Much of the focus on synaptotagmins has been in synaptic neurotransmitter release in neurons through their interactions
with soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNARE) (13, 32, 46, 49,
51, 52, 103). The general mechanism of this interaction begins with vesicular SNAREs interacting
with membrane SNAREs to dock the vesicle to the
membrane. Synaptotagmins bound to the vesicle
bind Ca2⫹ and are then able to interact with the
SNAREs to fuse the vesicle to the membrane and
expel the contents of the vesicle (FIGURE 2D).
Membrane repair is not solely dependent on synaptotagmins because studies using botulinum
toxin A, which cleaves SNARE proteins, showed
inhibition of resealing of sea urchin eggs (12). Disruption of the formation of the SNARE complex
using the cytoplasmic domain of synaptobrevin 2,
a SNARE protein, was also able to block membrane
resealing (125).
The relationship between synaptotagmins and
SNAREs is important because of their roles in Ca2⫹
sensing and vesicle membrane fusion. Syt I is
found exclusively in the nervous system (48, 138),
and antibodies against the C2A domain of Syt I
caused inhibition of membrane repair in severed
axons of squid and crayfish giant axons (40). This
same study used antibodies against a domain of a
SNARE protein, syntaxin, to inhibit interaction
with synaptotagmin’s C2A domain (40). Similar to
inhibition of Syt I, blocking syntaxin inhibited resealing of the severed axon (40). Syt VII is ubiquitously expressed and found proximal to the
membrane on lysosomal-associated membrane
protein 1 (LAMP-1)-positive lysosomes. Cells injured in the presence of Syt VII blocking antibodies
showed a decreased capacity to reseal membrane
disruptions (115), and fibroblasts taken from Syt
VII-deficient mice showed defective lysosomal exocytosis and decreased capacity to reseal their
plasma membrane (22).
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AHNAK, or desmoyokin, is a 629.1-kDa tripartite
nucleoprotein with potential functions as diverse
as fat metabolism, DNA repair, autoantigenicity,
cell signaling, calcium channel regulation, and tumor metastasis (33). It has been proposed that
interaction of AHNAK with annexin 2 and S100A10
regulates organization of the actin cytoskeleton
and architecture of the cell membrane, since
AHNAK-specific siRNA prevents actin cytoskeleton
reorganization (10). In membrane repair, AHNAK
has been associated with enlargeosomes, vesicles
that rapidly exocytose in response to calcium influx; however, its exact role within these vesicles is
unknown (14). AHNAK’s ability to bind actin may
signify a role for the protein in membrane resealing, although its presence within enlargeosomes
during recruitment to the membrane injury suggests earlier involvement in the repair process (33).
The co-localization of AHNAK with annexin 2
within vesicles is controversial, as studies also
have reported that annexin is on the cytosolic
face of the vesicle (89). In fact, calcium-sensitive
annexin 2 may be involved in recruitment of
AHNAK and S100A10 to the plasma membrane
in response to calcium-annexin binding (33).
Furthermore, co-immunoprecipitation and mass
spectrometry studies confirm that AHNAK interacts with dysferlin, which may localize and stabilize AHNAK at the sarcolemma through its
transmembrane domain, an interaction possibly
regulated by calpain 3 (67, 68) (FIGURE 2).
to the cell membrane, where the entire complex
acts as a scaffold for membrane repair (38, 81, 111,
118). Although S100A10 is known to be a central
player in the membrane repair complex, its exact
function in membrane repair has not been
identified. One model suggests that membrane
fusion may occur due to the ability of S100A10annexin A2 to bridge adjacent phospholipid
membranes (54).
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Conclusions
Eric X Beck provided helpful comments in the preparation of this manuscript.
This work was supported by the Muscular Dystrophy
Association, the OSU Physiology and Cell Biology Margaret T. Nishikawara Merit Scholarship Fund, and the National Institute of Arthritis and Musculoskeletal and Skin
Diseases of the National Institutes of Health under Award
No. AR-063084. The content is solely the responsibility of
the authors and does not necessarily represent the official
views of the National Institutes of Health.
Noah Weisleder is Founder and Chief Scientific Officer of
TRIM-edicine, a biotechnology company developing products targeting membrane repair, including rhMG53.
Author contributions: A.D.B., B.J.P., and N.W. prepared
figures; A.D.B., B.J.P., and N.W. drafted manuscript;
A.D.B., B.J.P., and N.W. edited and revised manuscript;
N.W. approved final version of manuscript.
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We have attempted to summarize the available
literature concerning the roles of many of the proteins known to contribute to the membrane repair
process. While clear evidence for the involvement
of the discussed proteins in membrane repair exists, it should be noted that other molecules, such
as Amphiphysin 2 (BIN1) (113), Lamp-1 (115), and
others, have been speculated to play a role in
membrane resealing due to the nature of their
actions in the cell or due to their interactions with
known members of the membrane repair machinery. For example, a recent study identified ATPase
EH-domain containing 2 as a novel membrane
repair protein that co-localizes at the site of membrane injury with F-actin and annexin A1 (91).
Moesin (membrane-organizing extension spike
protein) has been shown to interact with dysferlin
and appears to cross-link plasma membranes and
actin cytoskeletons (17). As more information
about specific proteins and membrane repair in
general is obtained, we expect that additional candidates will be discovered.
In considering the molecules known to be important in the membrane repair process, it becomes clear that many of the steps in membrane
repair are processes necessary for normal cellular
functions. For example, myosin, kinesin, annexin,
and SNAREs are involved in vesicle trafficking and
membrane fusion, integral events for numerous
cell activities (39). Calpain and affixin are important for cytoskeletal remodeling during cell motility, responses to the cell environment and mitosis
(66, 120). PTRF, ESCRT, ASM, and S100A10 are
active at the plasma membrane to form caveolae,
membrane buds, and other membrane invaginations as well as to mediate membrane fusion (17,
54, 74, 129). Dysferlin is integral to multiple activities, such as endocytosis, vesicle trafficking, membrane turnover, and others (31, 47, 58). Membrane
repair can be considered an emergency response
in which these cellular processes are used to reseal
the membrane and allow cell survival (57, 99, 105).
However, it is necessary to move beyond dissection
of this supportive cellular machinery to gain a full
understanding of membrane repair. Given the current understanding in the field, it is difficult to
define the prevailing membrane repair hypothesis,
and it is likely that there is no single mechanism
that is at work under all injury scenarios. Our current understanding of membrane repair is limited
to a subset of cellular functions and protein interactions, leaving compelling questions unanswered.
For example, how does the cell differentiate the
membrane repair process from normal cell functions and what are the specific signaling pathways
that allow this specific response to occur? Furthermore, what are the signals that direct the repair
machinery to the site of injury? While calciumdependent mechanisms are known signals for the
assembly of membrane repair proteins at the damage site and for fusion of membrane surfaces, additional mediators may exist. Discovery and
characterization of these mediators and pathways
are important next steps in understanding the
membrane repair process. 䡲
References
1.
Ampong BN, Imamura M, Matsumiya T, Yoshida M, Takeda S.
Intracellular localization of dysferlin and its association with
the dihydropyridine receptor. Acta Myol 24: 134 –144, 2005.
2.
Anderson LV, Davison K, Moss JA, Young C, Cullen MJ,
Walsh J, Johnson MA, Bashir R, Britton S, Keers S, Argov Z,
Mahjneh I, Fougerousse F, Beckmann JS, Bushby KM. Dysferlin is a plasma membrane protein and is expressed early in
human development. Hum Mol Genet 8: 855– 861, 1999.
3.
Babiychuk EB, Draeger A. Annexins in cell membrane dynamics. Ca2⫹-regulated association of lipid microdomains. J Cell
Biol 150: 1113–1124, 2000.
4.
Babiychuk EB, Monastyrskaya K, Potez S, Draeger A. Intracellular Ca2⫹ operates a switch between repair and lysis of
streptolysin O-perforated cells. Cell Death Differ 16: 1126 –
1134, 2009.
5.
Bansal D, Miyake K, Vogel SS, Groh S, Chen CC, Williamson
R, McNeil PL, Campbell KP. Defective membrane repair in
dysferlin-deficient muscular dystrophy. Nature 423: 168 –172,
2003.
6.
Bashir R, Britton S, Strachan T, Keers S, Vafiadaki E, Lako M,
Richard I, Marchand S, Bourg N, Argov Z, Sadeh M, Mahjneh
I, Marconi G, Passos-Bueno MR, Moreira EeS, Zatz M, Beckmann JS, Bushby K. A gene related to Caenorhabditis elegans spermatogenesis factor fer-1 is mutated in limb-girdle
muscular dystrophy type 2B. Nat Genet 20: 37– 42, 1998.
7.
Bazan NG, Marcheselli VL, Cole-Edwards K. Brain response
to injury and neurodegeneration: endogenous neuroprotective signaling. Ann NY Acad Sci 1053: 137–147, 2005.
8.
Beckmann JS, Spencer M. Calpain 3, the “gatekeeper” of
proper sarcomere assembly, turnover and maintenance.
Neuromuscul Disord 18: 913–921, 2008.
9.
Bement WM, Forscher P, Mooseker MS. A novel cytoskeletal
structure involved in purse string wound closure and cell
polarity maintenance. J Cell Biol 121: 565–578, 1993.
10. Benaud C, Gentil BJ, Assard N, Court M, Garin J, Delphin C,
Baudier J. AHNAK interaction with the annexin 2/S100A10
complex regulates cell membrane cytoarchitecture. J Cell
Biol 164: 133–144, 2004.
PHYSIOLOGY • Volume 30 • November 2015 • www.physiologyonline.org
445
REVIEWS
11. Bharadwaj A, Bydoun M, Holloway R, Waisman
D. Annexin A2 heterotetramer: structure and
function. Int J Mol Sci 14: 6259 – 6305, 2013.
12. Bi GQ, Alderton JM, Steinhardt RA. Calcium-regulated exocytosis is required for cell membrane
resealing. J Cell Biol 131: 1747–1758, 1995.
13. Bommert K, Charlton MP, DeBello WM, Chin GJ,
Betz H, Augustine GJ. Inhibition of neurotransmitter release by C2-domain peptides implicates
synaptotagmin in exocytosis. Nature 363: 163–
165, 1993.
14. Borgonovo B, Cocucci E, Racchetti G, Podini P,
Bachi A, Meldolesi J. Regulated exocytosis: a
novel, widely expressed system. Nat Cell Biol 4:
955–962, 2002.
15. Bouter A, Gounou C, Bérat R, Tan S, Gallois B,
Granier T, d’Estaintot BL, Pöschl E, Brachvogel B,
Brisson AR. Annexin-A5 assembled into two-dimensional arrays promotes cell membrane repair. Nat Commun 2: 270, 2011.
17. Cacciottolo M, Belcastro V, Laval S, Bushby K, di
Bernardo D, Nigro V. Reverse engineering gene
network identifies new dysferlin-interacting proteins. J Biol Chem 286: 5404 –5413, 2011.
18. Cai C, Masumiya H, Weisleder N, Matsuda N,
Nishi M, Hwang M, Ko JK, Lin P, Thornton A,
Zhao X, Pan Z, Komazaki S, Brotto M, Takeshima
H, Ma J. MG53 nucleates assembly of cell membrane repair machinery. Nat Cell Biol 11: 56 – 64,
2009.
19. Cai C, Masumiya H, Weisleder N, Pan Z, Nishi M,
Komazaki S, Takeshima H, Ma J. MG53 regulates
membrane budding and exocytosis in muscle
cells. J Biol Chem 284: 3314 –3322, 2009.
20. Cai C, Weisleder N, Ko JK, Komazaki S, Sunada
Y, Nishi M, Takeshima H, Ma J. Membrane repair
defects in muscular dystrophy are linked to altered interaction between MG53, caveolin-3, and
dysferlin. J Biol Chem 284: 15894 –15902, 2009.
21. Cao CM, Zhang Y, Weisleder N, Ferrante C,
Wang X, Lv F, Song R, Hwang M, Jin L, Guo J,
Peng W, Li G, Nishi M, Takeshima H, Ma J, Xiao
RP. MG53 constitutes a primary determinant of
cardiac ischemic preconditioning. Circulation
121: 2565–2574, 2010.
22. Chakrabarti S, Kobayashi KS, Flavell RA, Marks
CB, Miyake K, Liston DR, Fowler KT, Gorelick FS,
Andrews NW. Impaired membrane resealing and
autoimmune myositis in synaptotagmin VII-deficient mice. J Cell Biol 162: 543–549, 2003.
23. Chambers R, Chambers EL. Explorations into the
nature of the living cell. Acad Med 36: 966, 1961.
24. Chase TH, Cox GA, Burzenski L, Foreman O,
Shultz LD. Dysferlin deficiency and the development of cardiomyopathy in a mouse model of
limb-girdle muscular dystrophy 2B. Am J Pathol
175: 2299 –2308, 2009.
25. Chernomordik LV, Melikyan GB, Chizmadzhev
YA. Biomembrane fusion: a new concept derived
from model studies using two interacting planar
lipid bilayers. Biochim Biophys Acta 906: 309 –
352, 1987.
26. Cooper ST, Head SI. Membrane injury and repair
in the muscular dystrophies. Neuroscientist. In
press.
27. Cooper ST, Kizana E, Yates JD, Lo HP, Yang N,
Wu ZH, Alexander IE, North KN. Dystrophinopathy carrier determination and detection of protein deficiencies in muscular dystrophy using
lentiviral MyoD-forced myogenesis. Neuromuscul Disord 17: 276 –284, 2007.
446
29. Corrotte M, Almeida PE, Tam C, Castro-Gomes
T, Fernandes MC, Millis BA, Cortez M, Miller H,
Song W, Maugel TK, Andrews NW. Caveolae
internalization repairs wounded cells and muscle
fibers. Elife 2: e00926, 2013.
30. Corrotte M, Fernandes MC, Tam C, Andrews
NW. Toxin pores endocytosed during plasma
membrane repair traffic into the lumen of MVBs
for degradation. Traffic 13: 483– 494, 2012.
31. Covian-Nares JF, Koushik SV, Puhl HL, Vogel SS.
Membrane wounding triggers ATP release and
dysferlin-mediated intercellular calcium signaling. J Cell Sci 123: 1884 –1893, 2010.
32. Davis AF, Bai J, Fasshauer D, Wolowick MJ, Lewis
JL, Chapman ER. Kinetics of synaptotagmin responses to Ca2⫹ and assembly with the core
SNARE complex onto membranes. Neuron 24:
363–376, 1999.
33. Davis TA, Loos B, Engelbrecht AM. AHNAK: the
giant jack of all trades. Cell Signal 26: 2683–2693,
2014.
34. de Morrée A, Hensbergen PJ, van Haagen HH,
Dragan I, Deelder AM, ’t Hoen PA, Frants RR, van
der Maarel SM. Proteomic analysis of the dysferlin protein complex unveils its importance for
sarcolemmal maintenance and integrity. PLos
One 5: e13854, 2010.
35. Defour A, Van der Meulen JH, Bhat R, Bigot A,
Bashir R, Nagaraju K, Jaiswal JK. Dysferlin regulates cell membrane repair by facilitating injurytriggered acid sphingomyelinase secretion. Cell
Death Dis 5: e1306, 2014.
36. Demonbreun AR, Fahrenbach JP, Deveaux K,
Earley JU, Pytel P, McNally EM. Impaired muscle
growth and response to insulin-like growth factor
1 in dysferlin-mediated muscular dystrophy. Hum
Mol Genet 20: 779 –789, 2011.
37. Demonbreun AR, Lapidos KA, Heretis K, Levin S,
Dale R, Pytel P, Svensson EC, McNally EM.
Myoferlin regulation by NFAT in muscle injury,
regeneration and repair. J Cell Sci 123: 2413–
2422, 2010.
38. Dempsey BR, Rezvanpour A, Lee TW, Barber KR,
Junop MS, Shaw GS. Structure of an asymmetric
ternary protein complex provides insight for
membrane interaction. Structure 20: 1737–1745,
2012.
39. Derby MC, Gleeson PA. New insights into membrane trafficking and protein sorting. Intl Rev
Cytol 261: 47–116, 2007.
40. Detrait E, Eddleman CS, Yoo S, Fukuda M,
Nguyen MP, Bittner GD, Fishman HM. Axolemmal repair requires proteins that mediate synaptic vesicle fusion. J Neurobiol 44: 382–391, 2000.
41. Doherty KR, Cave A, Davis DB, Delmonte AJ,
Posey A, Earley JU, Hadhazy M, McNally EM.
Normal myoblast fusion requires myoferlin. Development 132: 5565–5575, 2005.
42. Draeger A, Babiychuk EB. Ceramide in plasma
membrane repair. Handb Exp Pharmacol 341–
353, 2013.
45. Duguez S, Bartoli M, Richard I. Calpain 3: a key
regulator of the sarcomere? FEBS J 273: 3427–
3436, 2006.
46. Earles CA, Bai J, Wang P, Chapman ER. The
tandem C2 domains of synaptotagmin contain
redundant Ca2⫹ binding sites that cooperate to
engage t-SNAREs and trigger exocytosis. J Cell
Biol 154: 1117–1123, 2001.
47. Evesson FJ, Peat RA, Lek A, Brilot F, Lo HP, Dale
RC, Parton RG, North KN, Cooper ST. Reduced
plasma membrane expression of dysferlin mutants is attributed to accelerated endocytosis via
a syntaxin-4-associated pathway. J Biol Chem
285: 28529 –28539, 2010.
48. Fox MA, Sanes JR. Synaptotagmin I and II are
present in distinct subsets of central synapses. J
Comp Neurol 503: 280 –296, 2007.
49. Fukuda M, Moreira JE, Lewis FM, Sugimori M,
Niinobe M, Mikoshiba K, Llinás R. Role of the C2B
domain of synaptotagmin in vesicular release and
recycling as determined by specific antibody injection into the squid giant synapse preterminal.
Proc Natl Acad Sci USA 92: 10708 –10712, 1995.
50. Gajic O, Lee J, Doerr CH, Berrios JC, Myers JL,
Hubmayr RD. Ventilator-induced cell wounding
and repair in the intact lung. Am J Respir Crit
Care Med 167: 1057–1063, 2003.
51. Geppert M, Goda Y, Hammer RE, Li C, Rosahl
TW, Stevens CF, Südhof TC. Synaptotagmin I: a
major Ca2⫹ sensor for transmitter release at a
central synapse. Cell 79: 717–727, 1994.
52. Geppert M, Goda Y, Stevens CF, Südhof TC. The
small GTP-binding protein Rab3A regulates a late
step in synaptic vesicle fusion. Nature 387: 810 –
814, 1997.
53. Gerke V, Creutz CE, Moss SE. Annexins: linking
Ca2⫹ signalling to membrane dynamics. Nat Rev
Mol Cell Biol 6: 449 – 461, 2005.
54. Gerke V, Moss SE. Annexins: from structure to
function. Physiol Rev 82: 331–371, 2002.
55. Ham YM, Mahoney SJ. Compensation of the AKT
signaling by ERK signaling in transgenic mice
hearts overexpressing TRIM72. Exp Cell Res 319:
1451–1462, 2013.
56. Han R, Bansal D, Miyake K, Muniz VP, Weiss RM,
McNeil PL, Campbell KP. Dysferlin-mediated
membrane repair protects the heart from stressinduced left ventricular injury. J Clin Invest 117:
1805–1813, 2007.
57. Han R, Campbell KP. Dysferlin and muscle membrane repair. Curr Opin Cell Biol 19: 409 – 416,
2007.
58. Han R, Frett EM, Levy JR, Rader EP, Lueck JD,
Bansal D, Moore SA, Ng R, Beltrán-Valero de
Bernabé D, Faulkner JA, Campbell KP. Genetic
ablation of complement C3 attenuates muscle
pathology in dysferlin-deficient mice. J Clin Invest 120: 4366 – 4374, 2010.
59. Heilbrunn LV. The Dynamics of Living Protoplasm. New York: Academic, 1956, p. 480 – 481.
60. Hernandez-Deviez DJ, Howes MT, Laval SH,
Bushby K, Hancock JF, Parton RG. Caveolin regulates endocytosis of the muscle repair protein,
dysferlin. J Biol Chem 283: 6476 – 6488, 2008.
43. Draeger A, Monastyrskaya K, Babiychuk EB.
Plasma membrane repair and cellular damage
control: the annexin survival kit. Biochem Pharmacol 81: 703–712, 2011.
61. Hernández-Deviez DJ, Martin S, Laval SH, Lo HP,
Cooper ST, North KN, Bushby K, Parton RG.
Aberrant dysferlin trafficking in cells lacking
caveolin or expressing dystrophy mutants of
caveolin-3. Hum Mol Genet 15: 129 –142, 2006.
44. Duann P, Li H, Lin P, Tan T, Wang Z, Chen K, Zhou
X, Gumpper K, Zhu H, Ludwig T, Mohler PJ,
Rovin B, Abraham WT, Zeng C, Ma J. MG53mediated cell membrane repair protects against
acute kidney injury. Sci Transl Med 7: 279ra236,
2015.
62. Hill MM, Bastiani M, Luetterforst R, Kirkham M,
Kirkham A, Nixon SJ, Walser P, Abankwa D,
Oorschot VM, Martin S, Hancock JF, Parton RG.
PTRF-Cavin, a conserved cytoplasmic protein required for caveola formation and function. Cell
132: 113–124, 2008.
PHYSIOLOGY • Volume 30 • November 2015 • www.physiologyonline.org
Downloaded from http://physiologyonline.physiology.org/ by 10.220.33.3 on June 15, 2017
16. Brose N, Petrenko AG, Südhof TC, Jahn R. Synaptotagmin: a calcium sensor on the synaptic
vesicle surface. Science 256: 1021–1025, 1992.
28. Corona BT, Garg K, Roe JL, Zhu H, Park KH, Ma
J, Walters TJ. Effect of recombinant human
MG53 protein on tourniquet-induced ischemiareperfusion injury in rat muscle. Muscle Nerve 49:
919 –921, 2014.
REVIEWS
63. Ho M, Post CM, Donahue LR, Lidov HG, Bronson
RT, Goolsby H, Watkins SC, Cox GA, Brown RH
Jr. Disruption of muscle membrane and phenotype divergence in two novel mouse models of
dysferlin deficiency. Hum Mol Genet 13: 1999 –
2010, 2004.
78. Kerr JP, Ziman AP, Mueller AL, Muriel JM, Kleinhans-Welte E, Gumerson JD, Vogel SS, Ward
CW, Roche JA, Bloch RJ. Dysferlin stabilizes
stress-induced Ca2⫹ signaling in the transverse
tubule membrane. Proc Natl Acad Sci USA 110:
20831–20836, 2013.
64. Holopainen JM, Angelova MI, Kinnunen PK. Vectorial budding of vesicles by asymmetrical enzymatic formation of ceramide in giant liposomes.
Biophys J 78: 830 – 838, 2000.
79. Keyel PA, Loultcheva L, Roth R, Salter RD, Watkins SC, Yokoyama WM, Heuser JE. Streptolysin
O clearance through sequestration into blebs
that bud passively from the plasma membrane. J
Cell Sci 124: 2414 –2423, 2011.
65. Holz RW, Hlubek MD, Sorensen SD, Fisher SK,
Balla T, Ozaki S, Prestwich GD, Stuenkel EL, Bittner MA. A pleckstrin homology domain specific
for phosphatidylinositol 4,5-bisphosphate (PtdIns-4,5-P2) and fused to green fluorescent protein identifies plasma membrane PtdIns-4,5-P2 as
being important in exocytosis. J Biol Chem 275:
17878 –17885, 2000.
67. Huang Y, de Morrée A, van Remoortere A,
Bushby K, Frants RR, den Dunnen JT, van der
Maarel SM. Calpain 3 is a modulator of the dysferlin protein complex in skeletal muscle. Hum
Mol Genet 17: 1855–1866, 2008.
81. Kobayashi K, Izawa T, Kuwamura M, Yamate J.
Dysferlin and animal models for dysferlinopathy.
J Toxicol Pathol 25: 135–147, 2012.
82. Kovtun O, Tillu VA, Ariotti N, Parton RG, Collins
BM. Cavin family proteins and the assembly of
caveolae. J Cell Sci 128: 1269 –1278, 2015.
83. Lee CS, Yi JS, Jung SY, Kim BW, Lee NR, Choo
HJ, Jang SY, Han J, Chi SG, Park M, Lee JH, Ko
YG. TRIM72 negatively regulates myogenesis via
targeting insulin receptor substrate-1. Cell Death
Differ 17: 1254 –1265, 2010.
68. Huang Y, Laval SH, van Remoortere A, Baudier J,
Benaud C, Anderson LV, Straub V, Deelder A,
Frants RR, den Dunnen JT, Bushby K, van der
Maarel SM. AHNAK, a novel component of the
dysferlin protein complex, redistributes to the
cytoplasm with dysferlin during skeletal muscle
regeneration. FASEB J 21: 732–742, 2007.
84. Lek A, Evesson FJ, Lemckert FA, Redpath GM,
Lueders AK, Turnbull L, Whitchurch CB, North
KN, Cooper ST. Calpains, cleaved mini-dysferlinC72, and L-type channels underpin calciumdependent muscle membrane repair. J Neurosci
33: 5085–5094, 2013.
69. Hwang M, Ko JK, Weisleder N, Takeshima H, Ma
J. Redox-dependent oligomerization through a
leucine zipper motif is essential for MG53-mediated cell membrane repair. Am J Physiol Cell
Physiol 301: C106 –C114, 2011.
85. Lennon NJ, Kho A, Bacskai BJ, Perlmutter SL,
Hyman BT, Brown RH Jr. Dysferlin interacts with
annexins A1 and A2 and mediates sarcolemmal
wound-healing. J Biol Chem 278: 50466 –50473,
2003.
70. Idone V, Tam C, Goss JW, Toomre D, Pypaert M,
Andrews NW. Repair of injured plasma membrane by rapid Ca2⫹-dependent endocytosis. J
Cell Biol 180: 905–914, 2008.
86. Lin P, Zhu H, Cai C, Wang X, Cao C, Xiao R, Pan
Z, Weisleder N, Takeshima H, Ma J. Nonmuscle
myosin IIA facilitates vesicle trafficking for
MG53-mediated cell membrane repair. FASEB J
26: 1875–1883, 2012.
71. Illa I, Serrano-Munuera C, Gallardo E, Lasa A,
Rojas-García R, Palmer J, Gallano P, Baiget M,
Matsuda C, Brown RH. Distal anterior compartment myopathy: a dysferlin mutation causing a
new muscular dystrophy phenotype. Ann Neurol
49: 130 –134, 2001.
72. Jaiswal JK, Marlow G, Summerill G, Mahjneh I,
Mueller S, Hill M, Miyake K, Haase H, Anderson
LV, Richard I, Kiuru-Enari S, McNeil PL, Simon
SM, Bashir R. Patients with a non-dysferlin Miyoshi myopathy have a novel membrane repair defect. Traffic 8: 77– 88, 2007.
73. Jia Y, Chen K, Lin P, Lieber G, Nishi M, Yan R,
Wang Z, Yao Y, Li Y, Whitson BA, Duann P, Li H,
Zhou X, Zhu H, Takeshima H, Hunter JC, McLeod
RL, Weisleder N, Zeng C, Ma J. Treatment of
acute lung injury by targeting MG53-mediated
cell membrane repair. Nat Commun 5: 4387,
2014.
74. Jimenez AJ, Maiuri P, Lafaurie-Janvore J, Divoux
S, Piel M, Perez F. ESCRT machinery is required
for plasma membrane repair. Science 343:
1247136, 2014.
75. Johnson CP, Chapman ER. Otoferlin is a calcium
sensor that directly regulates SNARE-mediated
membrane fusion. J Cell Biol 191: 187–197, 2010.
76. Jung SY, Ko YG. TRIM72, a novel negative feedback regulator of myogenesis, is transcriptionally
activated by the synergism of MyoD (or myogenin) and MEF2. Biochem Biophys Res Commun
396: 238 –245, 2010.
77. Kerr JP, Ward CW, Bloch RJ. Dysferlin at transverse tubules regulates Ca2⫹ homeostasis in
skeletal muscle. Front Physiol 5: 89, 2014.
87. Liu J, Aoki M, Illa I, Wu C, Fardeau M, Angelini C,
Serrano C, Urtizberea JA, Hentati F, Hamida MB,
Bohlega S, Culper EJ, Amato AA, Bossie K,
Oeltjen J, Bejaoui K, McKenna-Yasek D, Hosler
BA, Schurr E, Arahata K, de Jong PJ, Brown RH.
Dysferlin, a novel skeletal muscle gene, is mutated in Miyoshi myopathy and limb girdle muscular dystrophy. Nat Genet 20: 31–36, 1998.
88. Liu J, Zhu H, Zheng Y, Xu Z, Li L, Tan T, Park KH,
Hou J, Zhang C, Li D, Li R, Liu Z, Weisleder N, Zhu
D, Lin P, Ma J. Cardioprotection of recombinant
human MG53 protein in a porcine model of ischemia and reperfusion injury. J Mol Cell Cardiol
80: 10 –19, 2015.
89. Lorusso A, Covino C, Priori G, Bachi A, Meldolesi
J, Chieregatti E. Annexin2 coating the surface of
enlargeosomes is needed for their regulated exocytosis. EMBO J 25: 5443–5456, 2006.
90. Lostal W, Bartoli M, Roudaut C, Bourg N, Krahn
M, Pryadkina M, Borel P, Suel L, Roche JA, Stockholm D, Bloch RJ, Levy N, Bashir R, Richard I.
Lack of correlation between outcomes of membrane repair assay and correction of dystrophic
changes in experimental therapeutic strategy in
dysferlinopathy. PLos One 7: e38036, 2012.
91. Marg A, Schoewel V, Timmel T, Schulze A, Shah
C, Daumke O, Spuler S. Sarcolemmal repair is a
slow process and includes EHD2. Traffic 13:
1286 –1294, 2012.
92. Mariano A, Henning A, Han R. Dysferlin-deficient
muscular dystrophy and innate immune activation. FEBS J 280: 4165– 4176, 2013.
93. Matsuda C, Kameyama K, Suzuki A, Mishima W,
Yamaji S, Okamoto H, Nishino I, Hayashi YK.
Affixin activates Rac1 via betaPIX in C2C12 myoblast. FEBS Lett 582: 1189 –1196, 2008.
95. McNeil AK, Rescher U, Gerke V, McNeil PL. Requirement for annexin A1 in plasma membrane
repair. J Biol Chem 281: 35202–35207, 2006.
96. McNeil PL, Ito S. Gastrointestinal cell plasma
membrane wounding and resealing in vivo. Gastroenterology 96: 1238 –1248, 1989.
97. McNeil PL, Ito S. Molecular traffic through
plasma membrane disruptions of cells in vivo. J
Cell Sci 96: 549 –556, 1990.
98. McNeil PL, Khakee R. Disruptions of muscle fiber
plasma membranes. Role in exercise-induced
damage. Am J Pathol 140: 1097–1109, 1992.
99. McNeil PL, Kirchhausen T. An emergency response team for membrane repair. Nat Rev Mol
Cell Biol 6: 499 –505, 2005.
100. Mellgren RL, Miyake K, Kramerova I, Spencer MJ,
Bourg N, Bartoli M, Richard I, Greer PA, McNeil
PL. Calcium-dependent plasma membrane repair
requires m- or mu-calpain, but not calpain-3, the
proteasome, or caspases. Biochim Biophys Acta
1793: 1886 –1893, 2009.
101. Mellgren RL, Zhang W, Miyake K, McNeil PL.
Calpain is required for the rapid, calcium-dependent repair of wounded plasma membrane. J Biol
Chem 282: 2567–2575, 2007.
102. Micheva KD, Holz RW, Smith SJ. Regulation of
presynaptic phosphatidylinositol 4,5-biphosphate by neuronal activity. J Cell Biol 154: 355–
368, 2001.
103. Mikoshiba K, Fukuda M, Moreira JE, Lewis FM,
Sugimori M, Niinobe M, Llinás R. Role of the C2A
domain of synaptotagmin in transmitter release
as determined by specific antibody injection into
the squid giant synapse preterminal. Proc Natl
Acad Sci USA 92: 10703–10707, 1995.
104. Mishima W, Suzuki A, Yamaji S, Yoshimi R, Ueda
A, Kaneko T, Tanaka J, Miwa Y, Ohno S, Ishigatsubo Y. The first CH domain of affixin activates
Cdc42 and Rac1 through alphaPIX, a Cdc42/
Rac1-specific guanine nucleotide exchanging factor. Genes Cells 9: 193–204, 2004.
105. Miyake K, McNeil PL. Mechanical injury and repair of cells. Crit Care Med 31: S496 –S501, 2003.
106. Miyake K, McNeil PL. Vesicle accumulation and
exocytosis at sites of plasma membrane disruption. J Cell Biol 131: 1737–1745, 1995.
107. Miyake K, McNeil PL, Suzuki K, Tsunoda R, Sugai
N. An actin barrier to resealing. J Cell Sci 114:
3487–3494, 2001.
108. Nabhan JF, Hu R, Oh RS, Cohen SN, Lu Q. Formation and release of arrestin domain-containing
protein 1-mediated microvesicles (ARMMs) at
plasma membrane by recruitment of TSG101
protein. Proc Natl Acad Sci USA 109: 4146 –
4151, 2012.
109. Nagaraju K, Rawat R, Veszelovszky E, Thapliyal R,
Kesari A, Sparks S, Raben N, Plotz P, Hoffman
EP. Dysferlin deficiency enhances monocyte
phagocytosis: a model for the inflammatory onset of limb-girdle muscular dystrophy 2B. Am J
Pathol 172: 774 –785, 2008.
110. Ohsawa Y, Okada T, Kuga A, Hayashi S, Murakami T, Tsuchida K, Noji S, Sunada Y. Caveolin-3 regulates myostatin signaling. Mini-review.
Acta Myol 27: 19 –24, 2008.
111. Ozorowski G, Milton S, Luecke H. Structure of a
C-terminal AHNAK peptide in a 1:2:2 complex
with S100A10 and an acetylated N-terminal peptide of annexin A2. Acta Crystallogr D Biol Crystallogr 69: 92–104, 2013.
PHYSIOLOGY • Volume 30 • November 2015 • www.physiologyonline.org
447
Downloaded from http://physiologyonline.physiology.org/ by 10.220.33.3 on June 15, 2017
66. Honda S, Marumoto T, Hirota T, Nitta M, Arima
Y, Ogawa M, Saya H. Activation of m-calpain is
required for chromosome alignment on the
metaphase plate during mitosis. J Biol Chem 279:
10615–10623, 2004.
80. Kim SC, Kellett T, Wang S, Nishi M, Nagre N,
Zhou B, Flodby P, Shilo K, Ghadiali SN,
Takeshima H, Hubmayr RD, Zhao X. TRIM72 is
required for effective repair of alveolar epithelial
cell wounding. Am J Physiol Lung Cell Mol
Physiol 307: L449 –L459, 2014.
94. Matsuda C, Kameyama K, Tagawa K, Ogawa M,
Suzuki A, Yamaji S, Okamoto H, Nishino I,
Hayashi YK. Dysferlin interacts with affixin (betaparvin) at the sarcolemma. J Neuropathol Exp
Neurol 64: 334 –340, 2005.
REVIEWS
112. Parsegian VA, Rand RP, Gingell D. Lessons for
the study of membrane fusion from membrane
interactions in phospholipid systems. Ciba Found
Symp 103: 9 –27, 1984.
113. Prokic I, Cowling BS, Laporte J. Amphiphysin 2
(BIN1) in physiology and diseases. J Mol Med 92:
453– 463, 2014.
114. Raucher D, Sheetz MP. Characteristics of a membrane reservoir buffering membrane tension.
Biophys J 77: 1992–2002, 1999.
115. Reddy A, Caler EV, Andrews NW. Plasma membrane repair is mediated by Ca2⫹-regulated exocytosis of lysosomes. Cell 106: 157–169, 2001.
116. Redpath GM, Woolger N, Piper AK, Lemckert
FA, Lek A, Greer PA, North KN, Cooper ST.
Calpain cleavage within dysferlin exon 40a releases a synaptotagmin-like module for membrane repair. Mol Biol Cell 25: 3037–3048, 2014.
118. Rezvanpour A, Santamaria-Kisiel L, Shaw GS. The
S100A10-annexin A2 complex provides a novel
asymmetric platform for membrane repair. J Biol
Chem 286: 40174 – 40183, 2011.
119. Richard I, Roudaut C, Marchand S, Baghdiguian
S, Herasse M, Stockholm D, Ono Y, Suel L, Bourg
N, Sorimachi H, Lefranc G, Fardeau M, Sébille A,
Beckmann JS. Loss of calpain 3 proteolytic activity leads to muscular dystrophy and to apoptosisassociated IkappaBalpha/nuclear factor kappaB
pathway perturbation in mice. J Cell Biol 151:
1583–1590, 2000.
125. Shen SS, Tucker WC, Chapman ER, Steinhardt
RA. Molecular regulation of membrane resealing
in 3T3 fibroblasts. J Biol Chem 280: 1652–1660,
2005.
126. Steinhardt RA, Bi G, Alderton JM. Cell membrane resealing by a vesicular mechanism similar
to neurotransmitter release. Science 263: 390 –
393, 1994.
127. Sutton RB, Davletov BA, Berghuis AM, Südhof
TC, Sprang SR. Structure of the first C2 domain
of synaptotagmin I: a novel Ca2⫹/phospholipidbinding fold. Cell 80: 929 –938, 1995.
128. Swaggart KA, Demonbreun AR, Vo AH, Swanson
KE, Kim EY, Fahrenbach JP, Holley-Cuthrell J,
Eskin A, Chen Z, Squire K, Heydemann A, Palmer
AA, Nelson SF, McNally EM. Annexin A6 modifies muscular dystrophy by mediating sarcolemmal repair. Proc Natl Acad Sci USA 111: 6004 –
6009, 2014.
129. Tam C, Idone V, Devlin C, Fernandes MC, Flannery A, He X, Schuchman E, Tabas I, Andrews
NW. Exocytosis of acid sphingomyelinase by
wounded cells promotes endocytosis and plasma
membrane repair. J Cell Biol 189: 1027–1038,
2010.
130. Terasaki M, Miyake K, McNeil PL. Large plasma
membrane disruptions are rapidly resealed by
Ca2⫹-dependent vesicle-vesicle fusion events. J
Cell Biol 139: 63–74, 1997.
120. Rosenberger G, Gal A, Kutsche K. AlphaPIX associates with calpain 4, the small subunit of calpain, and has a dual role in integrin-mediated cell
spreading. J Biol Chem 280: 6879 – 6889, 2005.
131. Togo T, Steinhardt RA. Nonmuscle myosin IIA
and IIB have distinct functions in the exocytosisdependent process of cell membrane repair. Mol
Biol Cell 15: 688 – 695, 2004.
121. Roux I, Safieddine S, Nouvian R, Grati M, Simmler
MC, Bahloul A, Perfettini I, Le Gall M, Rostaing P,
Hamard G, Triller A, Avan P, Moser T, Petit C.
Otoferlin, defective in a human deafness form, is
essential for exocytosis at the auditory ribbon
synapse. Cell 127: 277–289, 2006.
132. Ueyama H, Kumamoto T, Nagao S, Masuda T,
Horinouchi H, Fujimoto S, Tsuda T. A new dysferlin gene mutation in two Japanese families
with limb-girdle muscular dystrophy 2B and Miyoshi myopathy. Neuromuscul Disord 11: 139 –
145, 2001.
122. Scheffer LL, Sreetama SC, Sharma N, Medikayala
S, Brown KJ, Defour A, Jaiswal JK. Mechanism of
Ca2⫹-triggered ESCRT assembly and regulation
of cell membrane repair. Nat Commun 5: 5646,
2014.
133. van der Kooi AJ, Frankhuizen WS, Barth PG,
Howeler CJ, Padberg GW, Spaans F, Wintzen
AR, Wokke JH, van Ommen GJ, de Visser M,
Bakker E, Ginjaar HB. Limb-girdle muscular dystrophy in the Netherlands: gene defect identified
in half the families. Neurology 68: 2125–2128,
2007.
123. Schiavo G, Gu QM, Prestwich GD, Söllner TH,
Rothman JE. Calcium-dependent switching of
the specificity of phosphoinositide binding to
synaptotagmin. Proc Natl Acad Sci USA 93:
13327–13332, 1996.
448
134. Waddell LB, Lemckert FA, Zheng XF, Tran J,
Evesson FJ, Hawkes JM, Lek A, Street NE, Lin P,
Clarke NF, Landstrom AP, Ackerman MJ,
Weisleder N, Ma J, North KN, Cooper ST. Dysferlin, annexin A1, and mitsugumin 53 are upregulated in muscular dystrophy and localize to
longitudinal tubules of the T-system with stretch.
J Neuropathol Exp Neurol 70: 302–313, 2011.
PHYSIOLOGY • Volume 30 • November 2015 • www.physiologyonline.org
135. Wang X, Xie W, Zhang Y, Lin P, Han L, Han P,
Wang Y, Chen Z, Ji G, Zheng M, Weisleder N,
Xiao RP, Takeshima H, Ma J, Cheng H. Cardioprotection of ischemia/reperfusion injury by cholesterol-dependent MG53-mediated membrane
repair. Circ Res 107: 76 – 83, 2010.
136. Weisleder N, Takeshima H, Ma J. Immuno-proteomic approach to excitation: contraction coupling in skeletal and cardiac muscle: molecular
insights revealed by the mitsugumins. Cell Calcium 43: 1– 8, 2008.
137. Weisleder N, Takizawa N, Lin P, Wang X, Cao C,
Zhang Y, Tan T, Ferrante C, Zhu H, Chen PJ, Yan
R, Sterling M, Zhao X, Hwang M, Takeshima M,
Cai C, Cheng H, Takeshima H, Xiao RP, Ma J.
Recombinant MG53 protein modulates therapeutic cell membrane repair in treatment of muscular dystrophy. Sci Transl Med 4: 139ra185,
2012.
138. Wendland B, Miller KG, Schilling J, Scheller RH.
Differential expression of the p65 gene family.
Neuron 6: 993–1007, 1991.
139. Wenzel K, Carl M, Perrot A, Zabojszcza J, Assadi
M, Ebeling M, Geier C, Robinson PN, Kress W,
Osterziel KJ, Spuler S. Novel sequence variants
in dysferlin-deficient muscular dystrophy leading
to mRNA decay and possible C2-domain misfolding. Hum Mutat 27: 599 – 600, 2006.
140. Wenzel K, Geier C, Qadri F, Hubner N, Schulz H,
Erdmann B, Gross V, Bauer D, Dechend R, Dietz
R, Osterziel KJ, Spuler S, Ozcelik C. Dysfunction
of dysferlin-deficient hearts. J Mol Med 85:
1203–1214, 2007.
141. Wenzel K, Zabojszcza J, Carl M, Taubert S, Lass
A, Harris CL, Ho M, Schulz H, Hummel O, Hubner
N, Osterziel KJ, Spuler S. Increased susceptibility
to complement attack due to down-regulation of
decay-accelerating factor/CD55 in dysferlin-deficient muscular dystrophy. J Immunol 175: 6219 –
6225, 2005.
142. Yamaji S, Suzuki A, Kanamori H, Mishima W, Yoshimi R, Takasaki H, Takabayashi M, Fujimaki K,
Fujisawa S, Ohno S, Ishigatsubo Y. Affixin interacts with alpha-actinin and mediates integrin signaling for reorganization of F-actin induced by
initial cell-substrate interaction. J Cell Biol 165:
539 –551, 2004.
143. Zhu H, Lin P, De G, Choi KH, Takeshima H,
Weisleder N, Ma J. Polymerase transcriptase release factor (PTRF) anchors MG53 protein to cell
injury site for initiation of membrane repair. J Biol
Chem 286: 12820 –12824, 2011.
Downloaded from http://physiologyonline.physiology.org/ by 10.220.33.3 on June 15, 2017
117. Rescher U, Gerke V. S100A10/p11: family,
friends and functions. Pflügers Arch 455: 575–
582, 2008.
124. Schissel SL, Schuchman EH, Williams KJ, Tabas I.
Zn2⫹-stimulated sphingomyelinase is secreted by
many cell types and is a product of the acid
sphingomyelinase gene. J Biol Chem 271: 18431–
18436, 1996.