A cluster of basic amino acid residues in the γ370-381 sequence of fibrinogen comprises a binding site for platelet integrin α IIbβ3 (glycoprotein IIb/IIIa)

Nataly Podolnikova, Oleg V. Gorkun, Ralph M. Loreth, Vivien C. Yee, Susan T. Lord, Tatiana Ugarova

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Adhesive interactions of platelet integrin αIIbβ 3 with fibrinogen and fibrin are central events in hemostasis and thrombosis. However, the mechanisms by which αIIbβ 3 binds these ligands remain incompletely understood. We have recently demonstrated that αIIbβ3 binds the γ365-383 sequence in the γC-domain of fibrin(ogen). This sequence contains neither the AGDV nor the RGD recognition motifs, known to bind αIIbβ3, suggesting the different specificity of the integrin. Here, using peptide arrays, mutant fibrinogens, and recombinant mutant γC-domains, we have examined the mechanism whereby αIIbβ3 binds γ365-383. The αIIbβ3-binding activity was localized within γ370-381, with two short sequences, γ370 ATWKTR 375 and γ376WYSMKK381, being able to independently bind the integrin. Furthermore, recognition of αIIbβ3 by γ370-381 depended on four basic residues, Lys373, Arg375, Lys380, and Lys 381. Simultaneous replacement of these amino acids and deletion of the γ408AGDV411 sequence in the recombinant γC-domain resulted in the loss of αIIbβ3- mediated platelet adhesion. Confirming the critical roles of the identified residues, abnormal fibrinogen Kaiserslautern, in which γLys380 is replaced by Asn, demonstrated delayed clot retraction and impaired αIIbβ3 binding. Also, a mutant recombinant fibrinogen modeled after the naturally occurring variant Osaka V (γArg375 → Gly) showed delayed clot retraction and reduced binding to purified αIIbβ3. These results identify the γ370-381 sequence of fibrin(ogen) as the binding site for αIIbβ3 involved in platelet adhesion and clot retraction and define the new recognition specificity of this integrin.

Original languageEnglish (US)
Pages (from-to)16920-16930
Number of pages11
JournalBiochemistry
Volume44
Issue number51
DOIs
StatePublished - Dec 27 2005
Externally publishedYes

Fingerprint

Basic Amino Acids
Platelet Glycoprotein GPIIb-IIIa Complex
Clot Retraction
Platelets
Integrins
Fibrinogen
Blood Platelets
Fibrin
Binding Sites
Abnormal Fibrinogens
Adhesion
Sequence Deletion
Hemostasis
Adhesives
Thrombosis
Ligands
Amino Acids
Peptides
estropipate

ASJC Scopus subject areas

  • Biochemistry

Cite this

A cluster of basic amino acid residues in the γ370-381 sequence of fibrinogen comprises a binding site for platelet integrin α IIbβ3 (glycoprotein IIb/IIIa). / Podolnikova, Nataly; Gorkun, Oleg V.; Loreth, Ralph M.; Yee, Vivien C.; Lord, Susan T.; Ugarova, Tatiana.

In: Biochemistry, Vol. 44, No. 51, 27.12.2005, p. 16920-16930.

Research output: Contribution to journalArticle

@article{4e26530564234440948e992149ad97ec,
title = "A cluster of basic amino acid residues in the γ370-381 sequence of fibrinogen comprises a binding site for platelet integrin α IIbβ3 (glycoprotein IIb/IIIa)",
abstract = "Adhesive interactions of platelet integrin αIIbβ 3 with fibrinogen and fibrin are central events in hemostasis and thrombosis. However, the mechanisms by which αIIbβ 3 binds these ligands remain incompletely understood. We have recently demonstrated that αIIbβ3 binds the γ365-383 sequence in the γC-domain of fibrin(ogen). This sequence contains neither the AGDV nor the RGD recognition motifs, known to bind αIIbβ3, suggesting the different specificity of the integrin. Here, using peptide arrays, mutant fibrinogens, and recombinant mutant γC-domains, we have examined the mechanism whereby αIIbβ3 binds γ365-383. The αIIbβ3-binding activity was localized within γ370-381, with two short sequences, γ370 ATWKTR 375 and γ376WYSMKK381, being able to independently bind the integrin. Furthermore, recognition of αIIbβ3 by γ370-381 depended on four basic residues, Lys373, Arg375, Lys380, and Lys 381. Simultaneous replacement of these amino acids and deletion of the γ408AGDV411 sequence in the recombinant γC-domain resulted in the loss of αIIbβ3- mediated platelet adhesion. Confirming the critical roles of the identified residues, abnormal fibrinogen Kaiserslautern, in which γLys380 is replaced by Asn, demonstrated delayed clot retraction and impaired αIIbβ3 binding. Also, a mutant recombinant fibrinogen modeled after the naturally occurring variant Osaka V (γArg375 → Gly) showed delayed clot retraction and reduced binding to purified αIIbβ3. These results identify the γ370-381 sequence of fibrin(ogen) as the binding site for αIIbβ3 involved in platelet adhesion and clot retraction and define the new recognition specificity of this integrin.",
author = "Nataly Podolnikova and Gorkun, {Oleg V.} and Loreth, {Ralph M.} and Yee, {Vivien C.} and Lord, {Susan T.} and Tatiana Ugarova",
year = "2005",
month = "12",
day = "27",
doi = "10.1021/bi051581d",
language = "English (US)",
volume = "44",
pages = "16920--16930",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "51",

}

TY - JOUR

T1 - A cluster of basic amino acid residues in the γ370-381 sequence of fibrinogen comprises a binding site for platelet integrin α IIbβ3 (glycoprotein IIb/IIIa)

AU - Podolnikova, Nataly

AU - Gorkun, Oleg V.

AU - Loreth, Ralph M.

AU - Yee, Vivien C.

AU - Lord, Susan T.

AU - Ugarova, Tatiana

PY - 2005/12/27

Y1 - 2005/12/27

N2 - Adhesive interactions of platelet integrin αIIbβ 3 with fibrinogen and fibrin are central events in hemostasis and thrombosis. However, the mechanisms by which αIIbβ 3 binds these ligands remain incompletely understood. We have recently demonstrated that αIIbβ3 binds the γ365-383 sequence in the γC-domain of fibrin(ogen). This sequence contains neither the AGDV nor the RGD recognition motifs, known to bind αIIbβ3, suggesting the different specificity of the integrin. Here, using peptide arrays, mutant fibrinogens, and recombinant mutant γC-domains, we have examined the mechanism whereby αIIbβ3 binds γ365-383. The αIIbβ3-binding activity was localized within γ370-381, with two short sequences, γ370 ATWKTR 375 and γ376WYSMKK381, being able to independently bind the integrin. Furthermore, recognition of αIIbβ3 by γ370-381 depended on four basic residues, Lys373, Arg375, Lys380, and Lys 381. Simultaneous replacement of these amino acids and deletion of the γ408AGDV411 sequence in the recombinant γC-domain resulted in the loss of αIIbβ3- mediated platelet adhesion. Confirming the critical roles of the identified residues, abnormal fibrinogen Kaiserslautern, in which γLys380 is replaced by Asn, demonstrated delayed clot retraction and impaired αIIbβ3 binding. Also, a mutant recombinant fibrinogen modeled after the naturally occurring variant Osaka V (γArg375 → Gly) showed delayed clot retraction and reduced binding to purified αIIbβ3. These results identify the γ370-381 sequence of fibrin(ogen) as the binding site for αIIbβ3 involved in platelet adhesion and clot retraction and define the new recognition specificity of this integrin.

AB - Adhesive interactions of platelet integrin αIIbβ 3 with fibrinogen and fibrin are central events in hemostasis and thrombosis. However, the mechanisms by which αIIbβ 3 binds these ligands remain incompletely understood. We have recently demonstrated that αIIbβ3 binds the γ365-383 sequence in the γC-domain of fibrin(ogen). This sequence contains neither the AGDV nor the RGD recognition motifs, known to bind αIIbβ3, suggesting the different specificity of the integrin. Here, using peptide arrays, mutant fibrinogens, and recombinant mutant γC-domains, we have examined the mechanism whereby αIIbβ3 binds γ365-383. The αIIbβ3-binding activity was localized within γ370-381, with two short sequences, γ370 ATWKTR 375 and γ376WYSMKK381, being able to independently bind the integrin. Furthermore, recognition of αIIbβ3 by γ370-381 depended on four basic residues, Lys373, Arg375, Lys380, and Lys 381. Simultaneous replacement of these amino acids and deletion of the γ408AGDV411 sequence in the recombinant γC-domain resulted in the loss of αIIbβ3- mediated platelet adhesion. Confirming the critical roles of the identified residues, abnormal fibrinogen Kaiserslautern, in which γLys380 is replaced by Asn, demonstrated delayed clot retraction and impaired αIIbβ3 binding. Also, a mutant recombinant fibrinogen modeled after the naturally occurring variant Osaka V (γArg375 → Gly) showed delayed clot retraction and reduced binding to purified αIIbβ3. These results identify the γ370-381 sequence of fibrin(ogen) as the binding site for αIIbβ3 involved in platelet adhesion and clot retraction and define the new recognition specificity of this integrin.

UR - http://www.scopus.com/inward/record.url?scp=29344472866&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=29344472866&partnerID=8YFLogxK

U2 - 10.1021/bi051581d

DO - 10.1021/bi051581d

M3 - Article

C2 - 16363805

AN - SCOPUS:29344472866

VL - 44

SP - 16920

EP - 16930

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 51

ER -