Soluble phosphatidylserine binds to a single identified site in the C2 domain of human factor Va

A. Srivastava; M. A. Quinn-Allen; S. W. Kim; W. H. Kane; B. R. Lentz

doi:10.1021/bi010449k

Soluble phosphatidylserine binds to a single identified site in the C2 domain of human factor V_a

A. Srivastava, M. A. Quinn-Allen, S. W. Kim, W. H. Kane, B. R. Lentz

Research output: Contribution to journal › Article › peer-review

26 Citations (Scopus)

Abstract

Factor V_a (FV_a) is a cofactor for the serine protease factor X_a that activates prothrombin to thrombin in the presence of Ca²⁺ and a membrane surface. FV_a is a heterodimer composed of one heavy chain (A1 and A2 domains) and one light chain (A3, C1, and C2 domains). We use fluorescence, circular dichroism, and equilibrium dialysis to demonstrate that (1) the FV C2 domain expressed in Sf9 cells binds one molecule of C6PS with a k_d of ∼2 μM, (2) stabilizing changes occur in the FV C2 domain upon C6PS binding, (3) the C6PS binding site in the FV C2 domain is located near residue Cys²¹¹³, which reacts with DTNB, and (4) binding to a PS-containing membrane is an order of magnitude tighter than that to soluble C6PS. Coupled with a recently published crystal structure of the C2 domain, these results support a model for the mechanism of C2-membrane interaction.

Original language	English
Pages (from-to)	8246-8255
Number of pages	10
Journal	Biochemistry
Volume	40
Issue number	28
DOIs	https://doi.org/10.1021/bi010449k
Publication status	Published - 2001 Jul 17
Externally published	Yes

ASJC Scopus subject areas

Biochemistry

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10.1021/bi010449k

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@article{108248f1e56b48d7991d23870ce969a3,

title = "Soluble phosphatidylserine binds to a single identified site in the C2 domain of human factor Va",

abstract = "Factor Va (FVa) is a cofactor for the serine protease factor Xa that activates prothrombin to thrombin in the presence of Ca2+ and a membrane surface. FVa is a heterodimer composed of one heavy chain (A1 and A2 domains) and one light chain (A3, C1, and C2 domains). We use fluorescence, circular dichroism, and equilibrium dialysis to demonstrate that (1) the FV C2 domain expressed in Sf9 cells binds one molecule of C6PS with a kd of ∼2 μM, (2) stabilizing changes occur in the FV C2 domain upon C6PS binding, (3) the C6PS binding site in the FV C2 domain is located near residue Cys2113, which reacts with DTNB, and (4) binding to a PS-containing membrane is an order of magnitude tighter than that to soluble C6PS. Coupled with a recently published crystal structure of the C2 domain, these results support a model for the mechanism of C2-membrane interaction.",

author = "A. Srivastava and Quinn-Allen, {M. A.} and Kim, {S. W.} and Kane, {W. H.} and Lentz, {B. R.}",

year = "2001",

month = jul,

day = "17",

doi = "10.1021/bi010449k",

language = "English",

volume = "40",

pages = "8246--8255",

journal = "Biochemistry",

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publisher = "American Chemical Society",

number = "28",

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T1 - Soluble phosphatidylserine binds to a single identified site in the C2 domain of human factor Va

AU - Srivastava, A.

AU - Quinn-Allen, M. A.

AU - Kim, S. W.

AU - Kane, W. H.

AU - Lentz, B. R.

PY - 2001/7/17

Y1 - 2001/7/17

N2 - Factor Va (FVa) is a cofactor for the serine protease factor Xa that activates prothrombin to thrombin in the presence of Ca2+ and a membrane surface. FVa is a heterodimer composed of one heavy chain (A1 and A2 domains) and one light chain (A3, C1, and C2 domains). We use fluorescence, circular dichroism, and equilibrium dialysis to demonstrate that (1) the FV C2 domain expressed in Sf9 cells binds one molecule of C6PS with a kd of ∼2 μM, (2) stabilizing changes occur in the FV C2 domain upon C6PS binding, (3) the C6PS binding site in the FV C2 domain is located near residue Cys2113, which reacts with DTNB, and (4) binding to a PS-containing membrane is an order of magnitude tighter than that to soluble C6PS. Coupled with a recently published crystal structure of the C2 domain, these results support a model for the mechanism of C2-membrane interaction.

AB - Factor Va (FVa) is a cofactor for the serine protease factor Xa that activates prothrombin to thrombin in the presence of Ca2+ and a membrane surface. FVa is a heterodimer composed of one heavy chain (A1 and A2 domains) and one light chain (A3, C1, and C2 domains). We use fluorescence, circular dichroism, and equilibrium dialysis to demonstrate that (1) the FV C2 domain expressed in Sf9 cells binds one molecule of C6PS with a kd of ∼2 μM, (2) stabilizing changes occur in the FV C2 domain upon C6PS binding, (3) the C6PS binding site in the FV C2 domain is located near residue Cys2113, which reacts with DTNB, and (4) binding to a PS-containing membrane is an order of magnitude tighter than that to soluble C6PS. Coupled with a recently published crystal structure of the C2 domain, these results support a model for the mechanism of C2-membrane interaction.

UR - http://www.scopus.com/inward/record.url?scp=0035902543&partnerID=8YFLogxK

U2 - 10.1021/bi010449k

DO - 10.1021/bi010449k

M3 - Article

C2 - 11444970

AN - SCOPUS:0035902543

SN - 0006-2960

VL - 40

SP - 8246

EP - 8255

JO - Biochemistry

JF - Biochemistry

IS - 28

ER -

Soluble phosphatidylserine binds to a single identified site in the C2 domain of human factor V_a

Abstract

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