Camp. Biochem.
Physiol. Vol. 118A, No. 4, pp. 1037-1044.
Copyright
0 1997 Elsevier Science Inc. All nghts reserved.
ISSN 0300-9629/97/$17.00
PI1 SOlOO-9629(97)0001~-1
1997
ELSEVIER
The Hemoglobin System of Pleuragramma anturcticum:
Correlation of Hematological and Biochemical
Adaptations With Life Style
Maurizjo Tamburrini ,* Rossanu D’ Amino,” Vito Carratore ,* Andreas Kunzmann , t
and Guido di Prisco”
*INSTITUTE
OF PROTEIN BIOCHEMISTRY AND ENZYMOLO~SY,C.N.R., VIA MARCONI 10, I-80125 NAPLES,
~INSTITUTE FOR POLAR Eco~ocu,
WISCHHOFSTRASSE 1-3, D-24148 KIEL, GERMAN)
ABSTRACT.
The hematological
untarcticum were investigated.
blooded antarctic
properties
determined.
The three hemoglobins
were differently
The complete
having three major hemoglobins,
amino acid sequence
showed strong Bohr and Root effects, and their oxygen-binding
regulated by temperature.
(or larval) remnants.
None of the three hemoglobins
Therefore,
this oxygen-transport
WORDS.
binding,
Amino
acid sequence,
Antarctic
fish, ecology,
of I’. antarcticurn can be considered
system is one of the most specialized ever
shelf seas are characterized
temperatures
contents
of more than 95% saturation
ditional
the Weddell
and advection
species
of this
among
(suborder
of the high-Antarctic
the more general
species
hematology,
and life
hemoglobin,
oxygen
Adin
system of currents
Notothenioidei,
processes
adaptations
necessary
notothenioids
is exceptional
It
of all notothenioids
for life in the water
the life history
(32)
fam(32).
target
of the high
for studies
The
biomass
whelmingly
significance
Antarctic,
in the pelagic
P. antarcticurn
on adaptation
logically
of pelagic
dominated
important
tion and migrates
and the only fully
shelf systems
systems
to extreme
is a suitable
environmental
conditions.
of water masses on
is the most abundant
with specializations
Because
production.
due to melting
distribution
antnrcticum
ily Nototheniidae)
combines
oxygen
fish life are found
a complex
stratification
and a specific
high
(33).
Pleurugramma
pelagic
governing
Sea, for example,
by low and
(29) and an extreme
and plankton
components
and ice drift, summer
the shelf
( - 1.6”C to -2.1”(Z),
in ice conditions
abiotic
evolution,
adaptation
mode of life and great biological
constant
seasonality
was
properties
thermodynamics
INTRODUCTION
The high-Antarctic
which
of the (Y- and /Q&~bin chains
found in fish. The data suggest a strong relationship between hematological/biochemical
style. COMP BIOCHEM PHYSIOL 118A;4:1037-1044,
1997. 0 1997 Elsevier Science Inc.
KEY
fish Pleurafframma
are at the lower end of the range of values known for red-
and of the suborder Notothenioidei
and fully characterized.
as evolutionary
system of the antarctic
ANI)
fish, suggesting a link with the sluggish mode of life of this species. P. antarcticurn is the only
species of the family Nototheniidae
were isolated
and the oxygen-transport
Most blood parameters
ITALY;
column.
and ecology
and due to its unique
P. antarcticurn
tion to water
species
in the midwater
ecosystem
der, this species
developed
reduction
ous and intermuscular
in vertebrates.
P.
growth
antarcticurn
has
distribu-
of notothenioid
adapta-
and may be the key species
and food web of the Antarctic
for the absence
neutral
of a swim blad-
buoyancy
by a combina-
and lipid deposition
in subcutane-
lipid sacs (15,21),
relatively
high
is fast. By the end of the second
ticum has accumulated
Address reprint requests to: G. dt Prisco, Institute
of Protein Biochemistry
and Enzynwlogy, C.N.R.. Via Marconi 10, I-80125 Naples, Italy. Tel. +3981-7257242;
Fax +39-81-5936689.
Presented at the 2nd Workshop
(Relationships
linking ecology, life style
and adaptwe evolution
m Antarcttc
hsh) of the European Science Foundation Netwxk
“Fishes of the Antarctic
Ocean,” Liege, September
21-23,
1995.
Recewed 22 April 1996; revised 25 November
1996; accepted 6 December 1996.
habitats
This eco-
water masses (1,31,32,36).
is the best example
column
Sea is over-
(24,33).
has a circum-Antarctic
across different
shelf (31). In compensation
tion of skeletal
fish of the Weddell
by P. antarcticurn
lipids, mainly
a unique
fecundity.
summer,
triacylglycerols,
strategy
Larval
P. anturcto pro-
vide buoyancy and also as energy stores (20). Conversely,
adults have an extremely
slow growth (23) and a very lowenergy-consuming
mode of life combined
with sluggish and
pelagic or benthopelagic
behavior
(16,22,35,36).
The fish respiratory system is an excellent
model for studies on environmental
adaptations
(45), which encompass
morphological,
physiological
and molecular
levels. Several
1038
M. Tamburrini
hematological
vironmental
parameters
Integration
studies
can be regulated
according
to en-
(50).
conditions
of hematology
of the oxygen-transport
fish. We report a thorough
MATERIALS
system of Antarctic
to the mode
(DE 52) was from Whatman,
from Cooper Biomedical,
Italy; Tris and bisTris from Sigma Chemical
St. Louis, MO, sequanal-grade
Co.,
reagents from Applied Bio-
systems, Foster City, CA, USA.
All other reagents were of
the highest purity commercially
available.
Fish were caught with Agassiz, bottom and benthopelagic
trawl during the PRV “Polarstem”
VII/4 (January-March
expeditions
EPOS Ant
1988, eastern and western Weddell
Sea) and Ant X/3 (March-May
1992, northeastern
dell Sea). Fish were immediately
transferred to aquaria and
allowed to recover for 12-24
16.6%
0.43 x lo’!/1
26.5 g/l
159.6 g/l
61.6 pg
4.32 ~01%
7.66
29.6 mm Hg
3.0 mm Hg
9.8
0.17
8.1
0.07
8.9
1.6
vein of 20 unanesthetized
hr.
specimens by means of heparin-
ized syringes. The blood parameters were investigated
mediately on board “Polarstern.”
im-
Samples were measured 3-
fold, and the results were averaged.
Blood gases (Paz and Pcol)
mM. Maximal
(100%)
oxygen saturation was de-
solutions. Meth-Hb
formation,
estimated from the absorbance at 630 nm, was always below
5-10%
of the
total.
change dH (kcal/mol;
heat of oxygen
The
overall
oxygenation
enthalpy
1 kcal = 4.184 kJ ), corrected for the
solubilization
(-3
kcal/mol),
was calcu-
lated by the integrated van? Hoff equation: dH = -4.574
[(CT1
TZ)/(Tl
- T2)] dlogP50/1000.
RESULTS
AND
Blood Parameters
Erythrocytes
direction)
DISCUSSION
were of similar size (11.5 pm in longitudinal
and shape (ellipsoid) as those of other nototheni-
ids.
Table
1 summarizes the values of the blood parameters
of P. antarcticurn, together with relevant statistical informa-
and pH were measured at
0°C with a modified Eschweiler (Kiel, Germany)
(38), the
number and Hb concen-
of Aethotatis mitopteryx, a closely related species of the same
cell counter.
were determined
lar Hb content
after the collection.
Hematocrit
and Hb concentration
by standard techniques
(MCH)
tration (MCHC)
The
with other nototheniids
erythrocyte
The number of red blood cells was counted with a Sysmex
immediately
ECO.
tion (37). In comparison
values of hematocrit,
tration are at the lower end of the range of values known
for red-blooded Antarctic fishes and very similar to those
CC-108
type 2031-02
blood-gassamples
were injected
2000,
0.05-0.1
termined in air-equilibrated
Wed-
blood samples were drawn from the caudal
analyzer System
Hematocrit
Erythrocytes
Hemoglobin
MCHC
MCH
Blood Q-CC
pH
PO2
PC02
Maidstone,
treated with t.-l-tosylamide-2-
phenylethylchloromethylketone,
Individual
SD
METHODS
UK; trypsin (EC 3.4.21.4),
Varese,
attention
in relation to the habitat.
AND
DEAE-cellulose
of
study on the oxygen-transport
system of P. antarcticurn, with special
of life and evolution
Value
and functional
insight into the principles that govern the development
adaptations
parameters of E antarcticurn
Parameter
with structural
(Hb) IS a useful approach to gaining
on hemoglobin
TABLE 1. Blood
et al.
(37). Mean cellu-
and mean corpuscular Hb concen-
were calculated according to Cobum
Fischer (7) and Hallmann
and
(28). From these values and the
subfamily. From the values of MCHC
and MCH and of oxy-
gen solubility in plasma (0.8 ~01%) (26), a total blood 02CC of 4.32 ~01% was calculated. The value ranges reported
for this ecologically
notothenioids
important parameter in other Antarctic
are 4.5-6.5
(42) and 3.3-7.7
(37). The val-
knowledge of oxygen solubility in plasma (0.8 ~01%) (26),
ues of PO* and Pcoz are likely to be affected by stress due
the total oxygen-carrying
to capture and handling. On the other hand, the difficulty
capacity (O&C)
was calculated.
Preparation of the hemolysates and cellulose acetate electrophoresis were carried out as described (8,9).
bin, tryptic digestions
and purification
In each glo-
of tryptic peptides
were carried out according to procedures previously described (10,48).
Amino acid sequencing was performed with an Applied
Biosystems (Foster City, CA, USA) automatic sequencer
model 477A, equipped with on-line detection of phenylthiohydantoin amino acids.
Oxygen-saturation
experiments were carried out at 2°C
as described
(9).
Oxygen-equilibrium
curves were deter-
mined tonometrically (9) at 2°C and 10°C in the pH range
6.5-8.0,
at a protein concentration,
on a heme basis, of
in keeping specimens
far prevented
Although
alive for long periods of time has so
to attempt recovery from stress.
it is difficult to establish an unambiguous corre-
lation between hematological parameters and life style, because we know only little about the mode of life or activity
of most Antarctic fish, the data reported here for P. untarcticum are clearly in agreement with the sluggish behavior
of this species.
Hb Multiplicity
and Molecular
Features
The hemolysate of notothenioids generally contains a single
major Hb (Hb 1) and often a minor component (Hb 2,
The Hemoglobins
approximately
chain
5% of the total).
in common
nodraco
mawsoni
(17-19),
major
cellulose.
two Hbs have
the p
of Cyg-
Hb 1 and Hb 2 share the a
phates
and chloride
in Table
analysis
indicated
(Hb
analysis
has
1, Hb 2 and Hb 3), which
by ion-exchange
HPLC
that P. anmrcticum
chromatography
of the purified
on DEAE-
Hbs showed
on the oxygen
affinity
Hb 1, Hb 2 and Hb 3, in the pH and temperature
and 2-10°C
effecters,
in the absence
show different
Some
implications
oxygenation
of these
enthalpies
functional
features
The Hb system of P. antarcticum
(49)) reported
the
tryptic
with
other
quences
known
chains
cleavage
have
The amino
Table
is built by two
of the four globins
in Fig. 1, were obtained
peptides
obtained
chemical
acid sequences
by the alignment
on the basis of sequence
Hb sequences
and by overlaps
with
the
or after
of Asp-Pro
terminus
(39).
The
acid residues,
a chains
of the
2 summarizes
N-terminus
bonds
142 and 146 amino
the degree
and of some
se-
IX and /3
ity major
components.
multiplicity
among
major
Hb
(13)]
with
differences
of the
Hbs and those
of the suborder
hance
cooperativity
species.
High
strong
alkaline
addressed
the other
components.
hand,
the chains
in common
with
Hb
of Hb 2 and Hb 3, which
1, showed
a high
identity
are
with
affinity
with
Hb 1 and Hb 3 show
50% cof Hb saturation)
though
low oxygen
the investigated
waters
gen-affinity
have
been
of Antarctic
by high
it lives
bodies
with
in the water
considerably
oxygen
column,
lower
for most
fish (37,41)
Hbs may be of advantage
cause
found
saturation,
living
high
for P. antarcticum,
where
oxygen
observed
zero in both);
in
servations
oxybe-
different
water
concentration
can
of oxygen
a very strong,
affinity
effector-enhanced,
on pH (alkaline
Bohr effect)
is regulated
en6.5 and
site even
should
differentiation
hy chloride
under
mainly
be
of the three
enthalpy
change
at
and organophosphates
decreased
in the
latter;
the
and absence
decrease
is
in contrast,
(especially
without
indicate
a
Hb 1 retains
effecters).
stronger
high oxyThese
ob-
effect
at
Bohr
physiological
temperatures
in Hb 1 (in the presence of effectors) and Hb 3 (in their absence).
In addition,
the moderate
effect
of temperature
on Hh 2 in the pH range
8.0 and on Hb 3 at pH 7.0 is indicative
of oxygen
loading
7.0-
of energy-saving
and unloading.
depen(46).
At 2°C the Bohr coefficient
(4 = Alog Pw/ApH)
ranges
from -0.9 to - 1.23. The Root effect (4) is also displayed
by all three Hbs, and its amplitude
than
lower (Fig. 4). A dramatic
enthalpy
clearly
than
pH in Hh 3 and also in Hb 2 (AH ap-
proaches
mechanisms
The Hhs display
is much
at lower
genation
easily occur.
dence
of effecters,
some
higher
of Hb 2, in the presence
of
ranges from 1.97 to 3.8 mm Hg. Alaffinities
species
characterized
is high.
to obtain
higher
a very strong
but drastically
required
but
deserve
affinities,
attention
to the thermodynamic
of oxygenation
fea-
effects),
Organophosphates
the hinding
However,
in the former,
of oxygen
see
by pH
also at pH 8.0 in Hb 3 only, indi-
interaction
heat
pressure
that
at all pH values
Antarctic
of Hb 1, Hb 2 and Hb 3 for oxygen
two;
oxygen-binding
and peculiarities
enhanced
The aftinity
regulated
in several
pH 8.0, further
At pH 8.0, P5p (partial
having
the Bohr and Root
those of minor Hbs of Antarctic
fish. The identity with globins of non-Antarctic
tish Hbs was lower, similar to other
fish.
highest
a single
newnesi,
notothenioids.
conditions.
identity was observed between the globin chains of P. antarcticum Hb 1 and those of major Hbs of Antarctic
fish. On
not
other
lower the oxygen
cating
the
have
generally
They have similar oxygen
those
non-Antarctic
which
(19).
Hbs are similar
be-
identity
affinity,
tures (e.g., they all display
comments.
is acetylated.
Trematomus
low oxygen
show
it displays
Notothenioidei,
[except
The three
is made of three high-aftin-
Therefore,
and organophosphates
they
respectively.
of sequence
tween the a and /? chains of I’. antarcticurn
of other Antarctic
fish, belonging to families
Notothenioidei,
of
homologies
directly
from
are ana-
that
mon. Thus,
of P. anturcticum
of the
(Fig. 4).
lyzed below.
Biochemical Adaptation and Life Style
the Hb system
ranges
and presence
Hb 1 has the a chain in common with Hb 2 and the p in
common with Hb 3. Hb 2 and Hb 3 have no chain in coma and two /3 chains.
The complete
amino
( P50) are reported
3.
of 7.0-8.0
components
were purified
The
with the only exception
(6), in which
chain.
Electrophoretic
three
1039
of P. antarcticurn
by the physi-
CONCLUDING
The respiratory
carrying
capacity
REMARKS
properties
AND PERSPECTIVES
of blood,
and oxygen
expecially
the oxygen-
affinity of Hb, respond
to evo-
ological effecters organophosphates
and chloride,
indicating a strong pH dependence
of Hb oxygenation
in air. Be-
lutionary
evolution
cause P. antarcticum,
as all Antarctic
notothenioids,
lacks
a swimbladder,
the presence of Root-effect
Hbs may be associated with the occurrence
of a choroid rete in the eye that,
gen-transport
system, such as reduction of erythrocyte
number, Hb concentration
and multiplicity
(19,51).
Pelagic P. antarcticurn
is the only species of the family
being poorly vascularized
in fish, depends on the diffusion
of oxygen from other tissues. The Bohr and Root effects in
Nototheniidae
and of the suborder Notothenioidei
having
three major Hbs, differently
regulated by temperature,
char-
the presence
of the effecters
respectively.
Additional
acterized by strong Bohr and Root effects.
very closely related benthopelagic
species
data
are illustrated
showing
the
in Figs 2 and 3,
effect
of phos-
selective
pressures.
In Antarctic
notothenioids,
has developed a variety of adaptations
in the oxy-
A. mitopteryx, a
(1) with a very
1040
M. Tamburrini
et al
aa
NA
A
AB
B
C
<______------_-, <--____________><_____>
CD
AC-~LSVKDKAAVKALWGKIGKS~AI~NDALSRMIWYPQTSK
1
25
EF
F
<___-___>
FG
G
<_--___--_---__--->
E
<_________--------->
50
GH
H
<----___------___--_>
HC
IPDLTAGLSDLSEQHAYKLRVDP~F~ILNHCILWIGIMFPKDFTPE~~LDKFLSAV~AERYR
75
100
125
cxb
NA
A
AB
B
C
<----__--------> <--_----__-----><---__>
CD
E
<---_----_-____---->
Ac-SLSTKDKEAVKAFWSKVSGHSEEIFNDALSRMLWYPQTKTYFSHWKEL~PGSAP~KHGM~GVGDAVSK
25
1
50
EF
F
<----___>
FG
G
<-____-___---__--->
GH
H
<----_----------__-->
HC
IPDLTAGLAELSELHAFTLRVDP~F~ILSHNILWFAIMFPNDFTPE~~VDKF~~KYR
125
75
100
pa
NA
C
B
A
<___-----_----><___-------_---><_____>
CD
D
E
<_----><----_-----__---_
yEWTANERAIITDIFSHIDYDDIG~~SRCLIVYPWTQRHFGAFGNLY~AEAIIGN~~GI~LHGLDR
25
50
1
H
G
GH
EF
F
FG
<_---__----__-----__>
<------->
<---___--_----____>
-->
GVKNMDNIAATYADLSVLHSQKLHVDPDNFVLLSDCITIVMAAKMGNAFT~IQGALQKFLAVWSALGKQYH
125
7;
lob
HC
Pb
D
E
C
CD
B
A
<_----><----------_____<___-----_----><______________><_____>
VEWTDFERATIADIFSKLEYDWGPATLARCLIVYPWTQRYFGSFGNLY~~IAEN~~HGTTIIHGLER
2j
50
i
NA
H
G
GH
FG
EF
F
<-_----____--_-___-_>
<------->
<--__----_----____>
-->
AVKNMDDITNTYAELSVLHSEKLINDPDNFKLLADCLTIWAARFGNAFTGHVQAAFQKFLAVWSALGRQYH
125
7;
100
HC
FIG. I. Amino acid sequences of the (Y and j3 chains of I? antarcticurn
Hhs. C? is identical in Hb 1 and Hb 2; (yb is the LYchain
of Hb 3. p” is identical in Hb 1 and Hb 3; fib is the p chain of Hb 2.
sluggish
mode of life, has a single Hb with a moderate
Bohr
effect and no Root effect (14). These observations
suggest
that these two species have to satisfy different oxygen demands, arising from special environmental
conditions.
In temperate
and tropical fish, ratios between
multiple
Hbs can vary seasonally and synthesis on demand is possible
(40). This may well be a typical
feature
of the oxygen-trans-
port system of P. antarcticurn. This species performs
migrations
(37)
through
water
masses
that
seasonal
(within
the
range typical of the environment)
may have different and
fluctuating
temperatures.
Thus, the main adaptive feature
of the Hb system of this fish should conceivably
be the response to the need to save energy during
gle Hb (or none at all) appears sufficient
migrations.
A sinto the other noto-
“Non-Antarctic
species.
/? chains
Notothenia coriiceps Hb 1, Hb 2
Tremotomus newnesi Hb 1, Hb 2
Pagothenia bernacchii Hb 1
Cygnodraco mawsoni Hb 1
Gymnodraco acuticeps
Pleuragromma antorcticum Hb 2
I’. antarcticurn Hb 1, Hb 3
Trematomus newnesi Hb C
Cygnodroco mawsoni Hb 2
Oncorhynchus mykiss* Hh I
0. mykiss* Hb IV
(43)
(2)
(12)
(13)
(5)
(6)
(48)
(49)
(49)
(13)
(6)
::;;
(49)
(11)
(13)
(3)
(44)
(10)
(13)
(5)
(6)
Reference
57
57
61
56
56
63
61
57
55
64
73
63
62
66
62
59
66
63
62
60
59
53
53
58
53
55
58
57
57
54
HbI
Hb IV
(27)
c.
65
64
66
67
65
86
67
89
Hb 2
mawsoni
0.
m yki..*
0.
myki.*
c.
61
63
65
64
65
90
67
93
Hb2
55
52
57
53
53
65
61
62
62
Hb I
T.
newnesi
57
62
62
62
62
66
64
63
58
60
Hb IV
0.
m yki..*
carpid
59
58
64
60
58
66
66
63
61
66
63
(30)
0.
myki..*
C.
car&P
identity (%) in (Yand /3 chains of some fish Hbs
(Ychains
Notothenia coriiceps Hb 1
Trematomw
newnrsi Hb I, Hb C
Pagothenia bermxchii Hb 1
Cygnodraco muwsoni Hb 1, Hb 2
Gymnodraco acuticeps
Pkuragrtxmmo antarcticurn Hb 3
P. antarcticurn Hb 1, Hb 2
Notothenia coriiceps Hb 2
Tremotomus newnesi Hb 2
Oncorhynchus mykiss* Hb I
0. mykiss* Hb IV
Species
TABLE 2. Sequence
N.
70
68
70
70
67
91
69
HbC
newnesi
T.
:;
92
70
63
66
70
Hb 2
car.
l?
86
84
90
82
80
72
Hb l,Hb3
antarc.
l?
95
89
92
87
85
72
Hb l,Hb2
antarc
P.
72
69
72
68
69
Hb 2
antarc.
I?
69
68
69
70
68
Hb3
antarc.
80
80
83
85
G.
acut.
82
92
91
93
G.
acut.
C.
88
84
87
Hb 1
mawsoni
c.
83
90
91
Hbl,HbZ
mawsoni
I?
90
93
Hbl
bem.
l?
89
97
Hbl
berm
T.
86
Hbl,HbZ
newnesi
T.
87
Hbl,HbC
newnesi
1042
M. Tamburrini
et al.
2.0
100
1.5
=:
80
e,
1.0
w
70
0
n
4
0.5
60
0.0
1
I
6.5
1
7.0
50
I
7.5
f
L
8.0
f
I
I
I
6.0
6.5
PH
instead
all sedentary
which
major
differ functionally
ation and influence
gen affinity.
optimal
ation
to speculate
and biochemical
savings
than
8.5
heat of oxygenregulation
of oxy-
has developed
suitable
to allow
extreme-environmental
in thermodynamic
in pH and organophosphate
FIG. 3. Oxygen saturation at atmospheric pressure as a function of pH of Hb 1 ( l ), Hb 2 (W) and Hb 3 (A). Experiments
were carried out in 100 mM Tris-HCI
or bisTris-HCI,
at
2”C, in the presence of 3 mM ATP. The value of 100% saturation was assigned to Hb at pH > 8.0.
can
the oxygenation-deoxygen-
Hbs differing
I
I
8.0
affinity,
that during evolution
adaptations
during
different-and
producing
rather
in overall
system of P. antarcticurn
cycle under
havior
mainly
It is tempting
energy
conditions,
Hbs with high oxygen
of pH on temperature
the oxygen-transport
physiological
dwellers. P. antarcticurn
bottom
rely on three
7.5
PH
FIG. 2. Oxygen-equilibrium
isotherms as a function of pH
of Hb 1 (O), Hb 2 (W) and Hb 3 (A). Experiments were carried out in 100 mM Tris-HCI or bisTris-HCl,
at 2”C, in the
presence of 100 mM NaCl and 3 mM ATP.
thenioids,
1
I
7.0
be-
regulation.
Hbs of P. antarcticl4m,
in the benthic
tionary
cance
unlike
(or larval)
distance
devoid
between
Hb 1 and the globins
the expression
remains
of multiple
genes
adult stage, in close similarity
of Hb 2
2). In this species,
with juveniles
refined
signifi-
data reveal high phy-
(Table
suggesting
found
as evolu-
of physiological
the sequence
and Hb 3 that are not in common
unpublished),
components
can be considered
remnants
(19), even though
logenetic
the minor
notothenioids,
mechanisms
high
also in the
(di Prisco et al.,
of regulation
The possibility that each component
(i.e., a fraction of the
total Hb) becomes selectively
functional
may explain the
within
need
relative
one of the most specialized ever found in fish, is remarkably
unique and appears designed to fit the unusual mode of life
(as well
of this fish through
for high
oxygen
affinity.
From this standpoint,
amounts
and on the basis of their
and of the awareness
as A. mitopteryx)
is a modem
that
species
I’. antarcticurn
(34), none of the three
the gene family.
Among
Notothenioidei,
adaptation
The
highly
this
refined
in the thermodynamic
three
oxygen-transport
molecular
system,
mechanisms
features
of each
Hbs may also serve different
purposes
of
Hb.
during
TABLE 3. Oxygen affinity ( IJsO)of the Hbs of F? antarcfi~um, at 2”C, in the absence and presence of the physiological effecters
Hb 1
Hb 2
Hb 3
PSO
100 mM NaCl
3mMATP
pH 6.5
pH 7.0
pH 7.5
pH 8.0
_
+
_
+
_
+
43.65
51.29
42.66
53.70
30.90
51.29
19.27
38.02
20.89
40.73
24.55
40.74
3.98
6.31
6.31
7.94
5.62
10.00
I.97
2.14
2.99
2.75
2.45
3.80
The Hemoglobins
1043
of P. antarcticurn
logical
able
16
activities
target
conformational
Hbs. These
12
studies
changes
investigations
on
the
P. antarcticurn
effect
in multisubunit
may shed
of
is a suit-
pressure
proteins
such
on
as
light
on this critical
and are a promising
area for future
This study is in the framework
of the Italian National Programme for
Antarctic Research. A. Kunmnn
thanks the German Science Foundation for financial support. G. di Prisco and M. Tambutini
are patef~l to the Alfred Wegener Institute. Bremerhawen.
Germany,
for the
invitation to participaate in the expedition Ant X/3 (March-May
1992),
in the northeastern
Weddell Sea. Thanks are due m the captain and
crew of the RV “Polarstern”
fur logistic support in the feld operations.
4
0
16
8
4
0
16
8
7.0
7.5
8.0
PH
FIG. 4. Heat of oxygenation of Hb 1, Hb 2 and Hb 3 (A, B
and C, respectively), in the absence (0, 0, A) and presence
(0, n, A) of 100 mM NaCl and 3 mM ATP. The values of
AH are corrected for the heat of solubilisation of oxygen in
water (-3 kcallmol).
ontogeny,
because young post-larvae,
juveniles
and adults
have different environmental
temperature
preferences
(32).
The
in fish. Therefore,
point of fish physiology
research.
8
(25);
for
depth range of P. antarcticurn
is from 0 to 900 m
hence, it is exposed to hydrostatic
pressure changes,
due to vertical displacement,
which may act as major selective factors (47), so far mostly overlooked,
regulating
bio-
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