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Deficiency
of Factor
X-Factor
V5 Binding
on the Platelets
of a Patient
With
a Bleeding
Disorder
By
Joseph
P. Miletich,
Nancy
William
Stanford,
H. Kane,
and
Factor
V (Vi) is essential
for binding
of
factor X to the surface
of platelets.
After
thrombin
treatment,
normal
platelets
release at least five times
more factor
V
activity
than is required
for maximal
factor
number
X,
factor
binding.
activity
The
concentration
obtained
after
of
factor
thrombin
of
in platelet-surface
V
X1 binding
and
prothrombin
activation
in normal
platelets;
some other components
limit the
W
E HAVE
RECENTLY
surface
per
platelet
ions,
more
ofstimulated
with
high
is reversible,
active
than
factor
proportional
reduced
the
binding
addition
normal
disorder
in
Kd
Xa.
*We
have
for
recently
the
Chemistry,
16634
Address
of
in part
from
Scientist,from
Submitted
Xa
C. M. Jackson,
Washington
Supported
Medical
factor
Division
May
reprint
decreased
of
that
normal
Majerus:
by Grants
the National
School
HLBI
the National
15, 1979;
requests
accepted
to Philip
form
J Biol
14147
July
Vol. 54, No. 5 (November),
1979
requires
calcium
Xa is 300,000-fold
prothrombin
and
stimulated
platelets
bovine
and
of General
plasma
platelets.
Large
of factor
V. rather
use the term,
St.
are
with
of factor
bleeding
submitted
of
Louis,
Center
by NIH
Medical
the
the
by
of
X,, binding
sites
disorder.6
The
is corrected
or obtained
excesses
than
factor
is
as well,
blocks
formation.
Platelets
activity
is associated
patient’s
by
as the
of factor
the procofactor
V,.
here
(W.
Va
V, is
H.
Kane.
for publication).
Internal
Medicine
and
Biological
Mo.
for
Research
Research
in
Service
Thrombosis)
Award
GM
and
07200,
Sciences.
5, 1979.
W. Majerus.
M.D..
Division
ton University
School
of Medicine.
Box 8125. 660 South
Euclid,
(c) 1979 by Grune
& Stratton,
Inc. 0006-4971/79/5405-0006$Ol.00/0
Blood,
factor
X,, to the
200 binding
sites
numbers
Chem,
(Specialized
of Health,
Institute
a
for-
following
stimulation
with
varying
degrees
Departments
of Medicine,
Institutes
suggests
thrombin
in solution
patients
we will
Hematology-Oncology.
University
factor
from
from
Therefore,
P. W.
Binding
patients’
an activated
binding.
coagulation
platelet-surface
Bound
thrombin
each
purified
stimulated
determined
platelet
M. J. Lindhout,
from
plasma
function
30 pM).
of thrombin
very little
have
V either
fraction
required
From
factor
to
platelet
III. Factor
V (V34
is essential
lgG specifically
and reversibly
severity
Xa to these
studies
moderate
patient’s
binding
of human
are approximately
activity
is found
Platelets
from
clinical
of factor
of
supernatant
HLBI
the
The
platelet
(apparent
V deficiency
to
binding.
with
and is specific
for factor
free factor
X,, in generating
binding
and the increased
rate
of factor
V in our studies.
Although
congenital
X
and
the
There
platelet,
significant
or other
agents.35
report
a
mation.
Xa
“resting”
thrombin
We
with
patient
protected
from inactivation
by antithrombin
since a homologous
monoclonal
anti-factor-V
factor
source
sites.
(MS.)
factors
described
platelets.’2
affinity
of binding
patient
factor
V activity
released
after thrombin
treatment
is normal,
but factor
X binding
is 20%-25%
of control values at saturation.
Abnormal
prothrombin
consumption
in a
stimula-
factor
a
L. Hofmann,
Majerus
severe
bleeding
abnormality
whose
platelets are deficient
in the
platelet-surface
component
required
for the factor
V-
tion of 1 7 normal platelets
is sufficient
to
allow half-maximal
factor X binding to
platelets
(10% normal,
90% factor-V
deficient).
Therefore,
factor
V activity
is not
limiting
Sandra
W.
Philip
Sites
of Hematology-Oncology.
St. Louis,
Mo.
Washing-
63110.
1015
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1016
MILETICH
activity
binding
factor
from either
sites beyond
Va S necessary
specific
platelet-surface
number
We
of these sources
do not increase
the total number
of factor
Xa
that normally
found with platelets
alone,
indicating
that while
for the platelet-factor-Xe
association,
it is not sufficient;
some
component
present
of prothrombin-converting
now reinforce
this concept
normal
human
necessary
platelets
at least
factor
We also describe
in small
binding.
factor
Xa
reduced
binding
Va
that
as much
factor
limits
after
the
stimulation
V activity
as is
Xa
binding,
to platelets
from a patient
with
et al.7), indicating
that a deficiency
Xafactor
ultimately
evidence
five times
site)
can
in the
presence
of normal
a moderately
severe
in the platelet-surface
Va,
factor
numbers
complexes.
by presenting
express
for maximal
factor
(Weiss
ET AL.
exist
and
excess
bleeding
disorder
component
(a
is associated
with
abnormal
hemostasis.
MATERIALS
AND
METHODS
Platelets
Platelet
visited
St.
control’s
isolation
was performed
Louis
for these studies,
(P.M.).
The
Experiments
patient
studied
MS.
on two
same
control
were
has been
occasions.
Louis (4-hr delay);
in the
conducted
reported
Both
times
her
On the second
in New
York,
within
in detail
et al.7 Factor
was drawn
(J.M.)
occasion,
The patient
EN.
obtained
routinely
in New
was drawn
blood
and an additional
has been described.’
at the same time
She
as the
6 hr of venipuncture.
by Weiss
blood
blood from the control
manner.
(W.K.)
as previously
reported.X
and her platelets
were
binding
and
from
(P.M.)
to her platelets
flown
simultaneously
was drawn
control
X
York
to St.
the first time and handled
her mother
sample
was
immediately
and
was drawn
father
as well
at the same
time
as a
in St.
Louis.
Factor-V-depleted
used
for
collected
was
all factor
Platelet
in the Tris-HCI
I0
by incubating
I0
platelets/mI
in the
phosphate
butler
2 U thrombin/ml
for 10 mm. The platelets
were
g for 5 mm and resuspended
in the phosphate buffer, and then the
mM
EDTA
use,
and
prior
to
bufler
used for binding
was determined
platelets/mI.
determined
since
platelets
We
platelet
it was
the
platelets
were
studies.
again
collected
This procedure
that
to
I09/ml
x i09 platelets/mI
(n
platelet
Factor
counts
in a microhematocrit
adjusted
the platelet
concentrations
possible
adjusted
had direct
by
centrifugation
and
appears to strip platelets
of
the
based
-
10). The
of 0.88
platelets
patient’s
x I09/ml
from
method
platelet
and 0.97
microscopy
differed
niicrohematocrit
assuming
a packed
ofsuspensions
by phase-contrast
patient’s
on the
centrifuge,
concentration
suspension
to I0
of all
normal
gave
platelet
in size.
phase
volume
suspensions
Suspensions
counts
on the two occasions
of
platelets/mi.
of 0.98
of
±
0.07
she was studied
x I09/ml.
Preparations
Human
prothrombin,
a gift
activity
5 mM
Just
concentration
also
was
prepared
2.5
V, activity.
1% equals
(SD)
were
with
at 2000
repeated.
resuspended
We
platelets
by centrifugation
process
used,
platelets
isolating
from
Dr.
was obtained
calcium
12,000
thrombin,
Craig
by incubating
chloride
g through
and
Jackson
and
factor
X were
2 mm.2
to the
The
for
supernatant
as previously
preparation
in the Tris-HCI
l0 platelets/mI
and 0.1 U thrombin/mI
oil for
prepared
was similar
buffer
10 mm. Platelets
fraction
reported.2
used
containing
Bovine
used for binding
were
removed
the
factor
V
earlier.6 Platelet factor V.
factor
studies
with
by centrifugation
Va activity
at
was
used
immediately.
Assays
Prothrombin,
were constructed
employing
the
X, were assayed
as reported
earlier.2
Thrombin
standard
B-3. Factor
V activity was determined
in a sensitive
purified reagents.6 Platelet factor V, activity
supernatant
reagent.
thrombin,
and factor
with
NIH
standard
from
platelets
stimulated
with
thrombin,
released
was measured
as described
above,
curves
assay
by immediately
diluting
into
V assay
the
factor
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
FACTOR
X,
-
Thrombin
described
V, BINDING
SITE DEFICIENCY
generation,
binding
of
1017
‘25l-factor
X,,
and
‘4C-serotonin
release
were
all
measured
as
previously.’2
RESULTS
Excess
Factor
V Activit’
specific
from
activity
either
The
factor
Va
factor
V molecules
assay activity
V in platelets
platelets
congenital
to
Front
Platelets
in coagulation
assays
and the
plasma
or platelets
is unknown.
associated
with
measurements.
was obtained
correct
absence
Normal
platelets
cannot
molecular
Thus,
the
be determined
weight
actual
from
of human
number
of
coagulation
However,
a minimal
estimate
of the quantity
by assaying
the ability
of thrombin-treated
specific
of factor
factor
X binding
V. The patient,
to
EN.,
platelets
from
a patient
was chosen
for these
of factor
normal
with
studies
because
there
is no specific
factor
X, binding
to her platelets
in the absence
of
added
factor
V.6 Preliminary
experiments
confirmed
that specific
factor
X, binding
and thrombin
formation
rates could
be restored
to normal
values
by the supernatant from thrombin-treated
control
platelets,
as had previously
been demonstrated
with
platelets
from
several
other
factor-V-deficient
patients.6
Maximal
specific
factor
X binding
to E.N ‘s platelets
was 1 .35 ng/
the maximum
specific
binding
to platelets
from
experiments
(1.30
ng/l08
platelets).
Since
stability
of the factor
V activity
released
X0
AND
BINDING
FACTOR
108 platelets,
the control
which
(P.M.)
detailed
information
from
platelets
under
TO MIXTURES
v-DEFICIENT
was similar
to
used in these
concerning
the
various
reaction
OF CONTROL
PLATELETS
U,
a)
4)
0
4)
0
C
z
0
a3
0
0.6
FRACTION
Fig.
Factor
X binding
concentration
to mixtures
V
-
DEFICIENT
0.8
PLATELETS
of control
(P.M.)
and factor-V-deficient
(EN.)
platelets.
platelets/mI,
while
the fraction
of factor-V-deficient
platelets
was increased
from 0 to 1. Reaction
mixtures
also initially contained
0.5 U thrombin/mI,
75
jg prothrombin/ml.
and 10 ng ‘25l-labeled
factor
X/ml
(specific
activity
2400
cpm/ng)
in buffer
containing
0.15 M sodium
chloride.
5 mM calcium
chloride,
5 mg bovine
serum
albumin/mI,
1 mg
glucose/mI.
and 20 mM Tris-HCI,
pH 7.4. Binding was determined
after 15 mm. Nonspecific
binding
was estimated
by addition
of excess
unlabeled
factor
X1 and subtracted.
Platelet
1.
FACTOR
was constant
at 10
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1018
MILETICH
conditions
mixtures
of
platelets,
constant,
and
but
as shown
Labeled
available,
we carried
and factor-V-deficient
than
both
initially
totally
the apparent
present
depleted
Kd and
at 75 tg/ml,
( I 5 mm).
for 2 mm before
the
platelets
was required
factor
obtained
amount
binding
indicates
of excess
could
that
factor
Xa
dissociation
since binding
control
constant
of factor
platelets,
from
l0
there
be at least
at
either
the amount
of excess
than these estimates.
least
factor
The
Factor
small
these
sites
sites
would
Xa-Factor
number
must
result
Recently,
Weiss
et al. have
and
glycoprotein
morphology)
mal serum
reported
composition
ultimately
rely on prothrombin
studied
the platelets
from this
The
patient’s
or both
where
such
normal
a patient
corrected
content,
her
platelet
parents,
release
V, and
the
activity
furthermore,
actually
be greater
and Methods,
described
above
per
platelet
that
abnormalities
plasma
suggested
that
of these
coagulation
(M.S.)
who
has
factors
a moderate
by platelet
transfusion.7
function,
phospholipid
as measured
conversion
patient,
her
‘4C-Serotonin
patient,
must
sites
postulated
in the
parents,
to
Extensive
composition,
by gel electrophoresis,
were within
normal
limits,
with the
prothrombin
times
and “platelet
factor
Thrombin-Induced
factor
sites.
However,
presence
of 10%
studies
would
be observed.
Such a defect
could only be
between
coagulation
factors
and platelets
is tested.
severe
bleeding
disorder
apparently
laboratory
studies
(including
platelet
protein
of
site,
.
concentration;
V binding
We
disorder
function
interaction
prothrombin
0.5 U throm-
Sites
properties.
in a bleeding
and routine
platelet
detected
when the
Prothrombin
before
with
V, as described
in Materials
platelets
in the experiments
Xafactor
unique
linearly.
five times
shown
in Fig. I The curve
surplus
factor
V. The exact
since we do not know the
“extra”
V or the affinity
Va Binding
of factor
have
bound).
from the platelet-surface
50% of maximal
in the
fivefold
of
Xa. If all of factor
V activity
factor
Xafactor
Va binding
a half-saturating
Normal
platelets
depleted
of factor
were substituted
for factor-V-deficient
with similar
results
(data
not shown).
Abnormal
amount
the dashed
line
platelets
contain
was not determined,
Va
using
Va binding
sites, was
increased
from 0 to 1,
was determined
was incubated
addition
of ‘251-factor
for saturation
of the
must
is
maximum
binding
mixture
(Kd) for factor
Xa is approximately
platelets/mi
of factor
XafaCtOr
platelets
was
the
and
Each
not exceed
the control
Va present
binding
assays
total
concentration
of factor
V necessarily
decreased
present
in excess
(approximately
.
bin/mi
control
out single-stage
platelets.
The
hence the total concentration
the fraction
of factor-V-deficient
in Fig. I The concentration
factor
X (‘251-factor
Xa)
was
greater
was
was
was not
control
ETAL.
size,
and
exception
of consistently
3 availabilities,”
both
abnorof which
presence
of platelets.
and several
controls.
We
have
Release
and
reaction
several
in
controls
were
response
concentrations
release
were:
of thrombin
in U/mI
necessary
MS.,
0.03 1; MS’s
father,
0.035;
P.M.,
0.020;
preparations
and J.M.,
0.045.
at a concentration
to
tested
thrombin
to
insure
was
for 50% of maximal
M.S.’s
mother,
0.043;
Maximal
release
of 0.5 U thrombin/mI.
was
obtained
with
that
normal.
the
The
‘4C-serotonin
W.K.,
0.021;
all
platelet
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FACTOR
X
-
V1 BINDING
Platelet-Accelerated
Platelets
Thrombin
were
tested
platelets
compared
occasion,
the
values
patient’s
(mother)
MS.
mixture
We
averaged
a
rate
of purified
factors
for
reported
values
On the
for
first
previously
(range,
4.3_7.2).6
of
U
1.8
thrombin/ml/min,
(J.M.)
value
of 5.5 U thrombin/ml/min.
supported
a rate of 2.0 U thrombin/mI/min,
(W.K.)
were
of ‘251-Factor
Platelets
reported5
platelets
MS’s
and
5.3
within
(P.M.)
the
On
U thrombin/mI/min.
normal
range
of this
were
values
X to Thrombin-Stimulated
also tested
for normal
at saturation
platelets
showed
several
the second
compared
to
Platelets
test:
from
4.6 (father)
and
4.3
251-factor
constants
occasion,
for their
ability
donors
were
I .65
a range
of 1.1-2.3
the time to reach
was
normal.
was decreased.
0.3 ng ‘25l-factor
Xa/108
platelets
However,
(MS.),
54 pM
the patient’s
parents
of 1.4 (father)
and
dissociation
constants
and
(W.K.),
were
1.1
to bind
‘251-Factor
Xa. Previously
0.40 ng factor
Xa bound
per I 08
±
ng. Preliminary
experiments
using
steady-state
‘251-factor
Xa binding
at
the
amount
of factor
Xa
bound
at
On the first occasion,
maximum
binding
to platelets
from
X,/l08
platelets,
as shown
in Fig. 2, compared
to 1.2 ng
for
the
in this experiment
were
values
obtained
(expressed
1.7 for W.K.,
Platelets
with
that
concentrations
saturation
M.S. was
pM
of 4.3
reaction
U thrombin/ml/min.
Binding
M.S.,
from
parents
Rates
thrombin
generation.
(SD)
U thrombin/ml/min
to a control
M.S.’s
platelets
control
Formation
in a standardized
their
ability
to enhance
normals
of 5.63 ± 0.92
occasion,
1019
SITE DEFICIENCY
1 .4 for
and
within
(mother)
of 46 pM
control
MS.,
(J.M.).
110
as ng
P.M.
The
42 pM
the
25
pM;
I-factor
apparent
(P.M.)
and
Xa/
51 pM
apparent
50
108
dissociation
pM. On the
platelets)
were
dissociation
in these
normal
range
ng ‘25I-factor
(father)
The
J.M.,
constants
experiments.
Since
0
4)
0
V
0
-
i/Xe
Free
(nq/mI)’
for
46
from
binding
apparent
we have
20
Fig. 2.
Double
reciprocal
plot of bound
versus free lThlfactor
X1 using
control
(J.M..
circles)
and
binding-site-deficient
(MS.,
triangles)
platelets.
Reaction
mixtures
and
nonspecific
binding
corrections
were
as in
Fig. 1, except
that ‘251-factor
X1 concentrations
were
varied
from
1 to 10 ng/ml.
Specific
activity
of the labeled
factor
X was
2600
cpm/ng.
0.4
were
Platelets
of values
with maximal
Xa/l08
platelets
and
(mother).
second
only
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
MILETICH
1020
been
able
to study
the
patient
twice,
factor
XaVa
receptor
sites have
is about
at the limit of a detectable
Platelet
Factor
The
activity
Va
(see
Materials
Control
(J.M.)
platelets
platelets
released
Localization
from
released
and
her platelets,
platelets
was
Defect
that
M.S.’s
platelets
was
measured
to the
factor
factor
platelets
V2.
When
Va containing
correct
the
these
UNTREATED
factor
and
0 ng
and
residual
in affinity
incubation
found
to be
platelets,
to platelets
V or some
soluble
with
normal.
while
from
factor-V-depleted
control
released
factor
platelets
‘251-factor
factor-Va-depleted
M.S.’s
FACTOR
from
not
from
thrombin-stimulated
of control
(W.K.)
stimulated
was
(W.K.)
platelets
deficient
bound
after
platelets
and
in her platelets.
1 .1 ng ‘2I-factor
platelets
Xa/l08
did
released
platelets
were tested
platelets
to rule out the
Va)
control
M.S.
inhibitor
supernatant
from her
factor
X binding
deficit
were
depletion
tested
of
in
the
V
DEPLETED
U
D
a)
0
o
following
Va/108
binding
Xa
factor
a)
4)
her
Surface
V. The patient’s
thrombin-stimulated
unidentified
before
U
factor
required
for normal
binding
(other
than
The results
are shown
in Fig. 3. Control
Xa/108
whether
platelets.
Platelet
of her
platelets
depleted
of factor
with the supernatant
from
some
0.56
Va/108
reduced
the factor
used
to
that
by
U factor
an abnormality
possibility
be certain
Methods)
released
0.75
of the
To demonstrate
result
cannot
V0 Activity
factor
thrombin
we
a normal
affinity.
The twofold
variation
difference
between
experiments.
ET AL.
D
NO ADDED
FACTOR
CONTROL
V
FACTOR
V
FACTOR
FROM
SITE-DEFICIENT
0.1
V
FROM
PLATELETS
BINDING
PLATELETS
.-%
a’
z
0
FACTOR 2
DEPLE TED
CONTROL
PLATELETS
BINDING
SITE-DEFICIENT
PLATELETS
Fig. 3.
Specific
‘25l-factor
X1 binding to control
(W.K.)
and binding-site-deficient
(M.S.)
platelets.
Reaction
mixtures
were as in Fig. 1. The specific
activity
of the 12S1.factor
X was 2000
cpm/ng.
Factor-V-depleted
platelets
and platelet
factor
V activities
were prepared
as described
in Materials
and Methods.
The factor
V activity
from 2 x lO platelets
was added to 10 platelets
in each case.
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
FACTORX1
V1BINDINGSITEDEFICIENCY
-
presence
of excess
factor
1021
Va activity
factor
Xa binding
was
platelets,
respectively).
derived
from
to the
the level
platelets,
observed
before
respectively).
In similar
factor
levels
range
found
with
experiments,
Xa binding
normal
factor
V. However,
neither
excess
binding
to these
depleted
rather,
(0.3
ng and
binding
0.2
bovine
was
restored
ng ‘251-factor
plasma
factor
of both control
and MS’s
factor-V-depleted
before
factor
V depletion
in each case.
observed
platelets,
251-faetor
Xa/ I 08
bound
0.3 before
.
platelets;
of purified
or M.S.’s
I 1 ng
from
MS.
of factor
increased
V depletion
addition
control
and
I
.
platelets
and 0.01 ng ‘251-factor
Xa after
being depleted
control
nor M.S.’s
platelet
factor
V activity
platelets
either
(1
restored
to normal
In this experiment,
to
Xa/108
V2 restored
platelets
only
to the
DISCUSSION
In previous
platelets
that
studies
we described
had undergone
the
the
release
binding
of
reaction.”2
factor
Factor
to the surface
of
was shown
to be
Xa
V
essential
for factor
X,, binding,
and it was suggested
that some unidentified
platelet
component
ultimately
limits
the number
of binding
sites.6
We now show
that
normal
platelets
release
at least
five times
more
factor
V than
is necessary
for
maximal
tion
factor
of factor
platelets/mi
l0
Xa binding
is
discovery
factor
normal
our
carried
of
sites
to
the
specific
binding
component
were
despite
Binding
sites
to be
phospholipid.
of factor
factor
congenital
thrombin
patient’s
parents
was
tested.
disorder
factor
formation
slightly
to
and
the
concentra-
stimulation
sites
on
from
of
108
correct
the
was
important
than
the
i’nherited
an autosomal
that are
that were
from our normal
range.6
of platelets
to accelerate
prothrombin
localizes
the
component
is
of the
even
normal
of
normal
in
200
if the
value
on
This
small
nanogram
of
values
at
all
is apparent.
The association
of factor
Xa, as with patients
in normal
an
this
were
30%-35%.
generation
per
range
underscores
event
for
from
analysis,
20%-25%
for this finding
platelet
binding
to define
not excluded
X2 binding
by values
the
the
required
fraction
phospholipid
content
a defect
in less than
phospholipid
formation
rates
rate of thrombin
higher
of
of
indicates
abnormality
nature
to
number
clearly
platelet
factor
of the supernatant
chemical
MS.
a reduced
V2 activity
in total
V deficiency,
values
for factor
be characterized
be distinguishable
The ability
factor
The
No explanation
with decreased
parents
in an attempt
were normal,
we have
patient
has
state would
system,
thrombin
binding
disorder
unidentified
The failure
reflected
thrombin
as the
is an
in vitro
Weiss et al.7 found normal
be unreasonable
to expect
Xa to platelets
bound
Xa
yet
surface.
Although
it would
occasions,
while
was consistent,
concentrations
of a bleeding
normal
platelets
platelet
several
difference
in our
solution
following
to half-saturate
a bleeding
some
specific
as
thrombin
formation.
unknown
at this time.
the patient’s
platelets,
with
with
thrombin-stimulated
component
at least
earlier
studies
on the kinetics
of factor
X,, binding
out under
conditions
of saturating
factor
V.
of a patient
X2 binding
involvement
platelet-surface
that,
released
into
least
sufficient
at
platelets/mI.
Therefore,
normal
platelets
were
The
and
Va activity
fact
that
platelet-surface
disorder.
recessive
Even though
trait.
Since
the
the
the heterozygous
which
would
not
hemostasis.
25% of normal,
75% of normal,
activation
We
has
studied
been
the
termed
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
MILETICH
1022
“platelet
necessarily
factor
3” (PF-3).
Tests
detect
abnormalities
abnormality
Thus,
of other
while
studied
had
platelet-dependent
patient
with
M.S.
normal
platelet
and
This
also
patient
tooth
extractions
trauma
not
PF-3
factor
reactions
test
(R.G.)
Xa, binding
a normal
could
had
a mild
2 occasions
but
lead
no residual
platelets
disorder
with
increased
transfusions.
We
abnormalities
of platelet
prothrombin
Residual
prothrombin
in serum
despite
that
that
with
a normal
hemorrhage
bleeding
the
best
after
after
test
mild
to
activation
is a serum
prothrombin
normal
plasma
coagulation
factors
prothrombin
we
in serum
U thrombin/mI/min.
of increased
suggest
not
since
results.
patient
prothrombin
of 4.5
a history
do
to abnormal
rate
and
clinically
activation,
test,9 another
generation
bleeding
in platelet-surface
PF-3
1 ng X bound/108
thrombin
on
requiring
an abnormality
this parameter
prothrombin
did have an abnormal
an abnormal
Kd of 57 pM
used to measure
of platelet-surface
ET AL.
detect
time.7
suggests
activation.
ACKNOWLEDGMENT
The
authors
thank
Dr.
JP,
Jackson
Weiss
Harvey
for providing
access
to the
patient
MS.,
her parents,
as well
as
patient R.G.
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1. Miletich
Interaction
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4036,
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Majerus
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The
detection
with
mild
bleeding
32:872-883,
1962
by
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
1979 54: 1015-1022
Deficiency of factor Xa-factor Va binding sites on the platelets of a patient
with a bleeding disorder
JP Miletich, WH Kane, SL Hofmann, N Stanford and PW Majerus
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