Fibrinogen binds to integrin α5β1 via the Carboxyl-terminal RGD site of the Aα-chain

K. Suehiro, J. Mizuguchi, K. Nishiyama, S. Iwanaga, David Farrell, S. Ohtaki

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Fibrinogen interactions with vascular endothelial cells are implicated in various physiological and pathophysiological events, including angiogenesis and wound healing. We have shown previously that integrin α5β1 is a fibrinogen receptor on endothelial cells [Suehiro, K., Gailit, J., and Plow, E.F. (1997) J. Biol. Chem. 272, 5360-5366]. In the present study, we have characterized fibrinogen interactions with purified α5β1 and have identifled the recognition sequence in fibrinogen for α5β1. The binding of fibrinogen to immobilized α5β1 was selectively supported by Mn2+. Fibrinogen bound to purified α5β1 in a time-dependent, specific, and saturable manner in the presence of Mn2+, and the binding was blocked completely by Arg-Gly-Asp (RGD)-containing peptides and by anti-α5 and anti-α5β1 monoclonal antibodies. A monoclonal antibody directed to the C-termlnal RGD sequence at Aα572-574 significantly inhibited the binding of fibrinogen to α5β1, whereas monoclonal antibodies directed to either the N-terminal RGD sequence at Aα95-97 or the C-terminus of the γ-chain did not. Furthermore, substituting RGE for RGD at position Aα95-97 in recombinant fibrinogen had a minimal effect on binding, whereas substituting RGE for RGD at position Aα572-574 decreased binding by 90%. These results demonstrate that the C-terminal RGD sequence at Aα572-574 is required for the interaction of fibrinogen with α5β1.

Original languageEnglish (US)
Pages (from-to)705-710
Number of pages6
JournalJournal of Biochemistry
Volume128
Issue number4
StatePublished - 2000

Fingerprint

Integrins
Fibrinogen
Monoclonal Antibodies
Endothelial cells
Endothelial Cells
Fibrinogen Receptors
Wound Healing
Peptides

Keywords

  • Divalent cations
  • Fibrinogen
  • Fibronectin receptor
  • Integrin
  • RGD

ASJC Scopus subject areas

  • Biochemistry

Cite this

Suehiro, K., Mizuguchi, J., Nishiyama, K., Iwanaga, S., Farrell, D., & Ohtaki, S. (2000). Fibrinogen binds to integrin α5β1 via the Carboxyl-terminal RGD site of the Aα-chain. Journal of Biochemistry, 128(4), 705-710.

Fibrinogen binds to integrin α5β1 via the Carboxyl-terminal RGD site of the Aα-chain. / Suehiro, K.; Mizuguchi, J.; Nishiyama, K.; Iwanaga, S.; Farrell, David; Ohtaki, S.

In: Journal of Biochemistry, Vol. 128, No. 4, 2000, p. 705-710.

Research output: Contribution to journalArticle

Suehiro, K, Mizuguchi, J, Nishiyama, K, Iwanaga, S, Farrell, D & Ohtaki, S 2000, 'Fibrinogen binds to integrin α5β1 via the Carboxyl-terminal RGD site of the Aα-chain', Journal of Biochemistry, vol. 128, no. 4, pp. 705-710.
Suehiro K, Mizuguchi J, Nishiyama K, Iwanaga S, Farrell D, Ohtaki S. Fibrinogen binds to integrin α5β1 via the Carboxyl-terminal RGD site of the Aα-chain. Journal of Biochemistry. 2000;128(4):705-710.
Suehiro, K. ; Mizuguchi, J. ; Nishiyama, K. ; Iwanaga, S. ; Farrell, David ; Ohtaki, S. / Fibrinogen binds to integrin α5β1 via the Carboxyl-terminal RGD site of the Aα-chain. In: Journal of Biochemistry. 2000 ; Vol. 128, No. 4. pp. 705-710.
@article{890fd47f21f34bcc945e9b11f5f6b224,
title = "Fibrinogen binds to integrin α5β1 via the Carboxyl-terminal RGD site of the Aα-chain",
abstract = "Fibrinogen interactions with vascular endothelial cells are implicated in various physiological and pathophysiological events, including angiogenesis and wound healing. We have shown previously that integrin α5β1 is a fibrinogen receptor on endothelial cells [Suehiro, K., Gailit, J., and Plow, E.F. (1997) J. Biol. Chem. 272, 5360-5366]. In the present study, we have characterized fibrinogen interactions with purified α5β1 and have identifled the recognition sequence in fibrinogen for α5β1. The binding of fibrinogen to immobilized α5β1 was selectively supported by Mn2+. Fibrinogen bound to purified α5β1 in a time-dependent, specific, and saturable manner in the presence of Mn2+, and the binding was blocked completely by Arg-Gly-Asp (RGD)-containing peptides and by anti-α5 and anti-α5β1 monoclonal antibodies. A monoclonal antibody directed to the C-termlnal RGD sequence at Aα572-574 significantly inhibited the binding of fibrinogen to α5β1, whereas monoclonal antibodies directed to either the N-terminal RGD sequence at Aα95-97 or the C-terminus of the γ-chain did not. Furthermore, substituting RGE for RGD at position Aα95-97 in recombinant fibrinogen had a minimal effect on binding, whereas substituting RGE for RGD at position Aα572-574 decreased binding by 90{\%}. These results demonstrate that the C-terminal RGD sequence at Aα572-574 is required for the interaction of fibrinogen with α5β1.",
keywords = "Divalent cations, Fibrinogen, Fibronectin receptor, Integrin, RGD",
author = "K. Suehiro and J. Mizuguchi and K. Nishiyama and S. Iwanaga and David Farrell and S. Ohtaki",
year = "2000",
language = "English (US)",
volume = "128",
pages = "705--710",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "4",

}

TY - JOUR

T1 - Fibrinogen binds to integrin α5β1 via the Carboxyl-terminal RGD site of the Aα-chain

AU - Suehiro, K.

AU - Mizuguchi, J.

AU - Nishiyama, K.

AU - Iwanaga, S.

AU - Farrell, David

AU - Ohtaki, S.

PY - 2000

Y1 - 2000

N2 - Fibrinogen interactions with vascular endothelial cells are implicated in various physiological and pathophysiological events, including angiogenesis and wound healing. We have shown previously that integrin α5β1 is a fibrinogen receptor on endothelial cells [Suehiro, K., Gailit, J., and Plow, E.F. (1997) J. Biol. Chem. 272, 5360-5366]. In the present study, we have characterized fibrinogen interactions with purified α5β1 and have identifled the recognition sequence in fibrinogen for α5β1. The binding of fibrinogen to immobilized α5β1 was selectively supported by Mn2+. Fibrinogen bound to purified α5β1 in a time-dependent, specific, and saturable manner in the presence of Mn2+, and the binding was blocked completely by Arg-Gly-Asp (RGD)-containing peptides and by anti-α5 and anti-α5β1 monoclonal antibodies. A monoclonal antibody directed to the C-termlnal RGD sequence at Aα572-574 significantly inhibited the binding of fibrinogen to α5β1, whereas monoclonal antibodies directed to either the N-terminal RGD sequence at Aα95-97 or the C-terminus of the γ-chain did not. Furthermore, substituting RGE for RGD at position Aα95-97 in recombinant fibrinogen had a minimal effect on binding, whereas substituting RGE for RGD at position Aα572-574 decreased binding by 90%. These results demonstrate that the C-terminal RGD sequence at Aα572-574 is required for the interaction of fibrinogen with α5β1.

AB - Fibrinogen interactions with vascular endothelial cells are implicated in various physiological and pathophysiological events, including angiogenesis and wound healing. We have shown previously that integrin α5β1 is a fibrinogen receptor on endothelial cells [Suehiro, K., Gailit, J., and Plow, E.F. (1997) J. Biol. Chem. 272, 5360-5366]. In the present study, we have characterized fibrinogen interactions with purified α5β1 and have identifled the recognition sequence in fibrinogen for α5β1. The binding of fibrinogen to immobilized α5β1 was selectively supported by Mn2+. Fibrinogen bound to purified α5β1 in a time-dependent, specific, and saturable manner in the presence of Mn2+, and the binding was blocked completely by Arg-Gly-Asp (RGD)-containing peptides and by anti-α5 and anti-α5β1 monoclonal antibodies. A monoclonal antibody directed to the C-termlnal RGD sequence at Aα572-574 significantly inhibited the binding of fibrinogen to α5β1, whereas monoclonal antibodies directed to either the N-terminal RGD sequence at Aα95-97 or the C-terminus of the γ-chain did not. Furthermore, substituting RGE for RGD at position Aα95-97 in recombinant fibrinogen had a minimal effect on binding, whereas substituting RGE for RGD at position Aα572-574 decreased binding by 90%. These results demonstrate that the C-terminal RGD sequence at Aα572-574 is required for the interaction of fibrinogen with α5β1.

KW - Divalent cations

KW - Fibrinogen

KW - Fibronectin receptor

KW - Integrin

KW - RGD

UR - http://www.scopus.com/inward/record.url?scp=0033772764&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033772764&partnerID=8YFLogxK

M3 - Article

C2 - 11011154

AN - SCOPUS:0033772764

VL - 128

SP - 705

EP - 710

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 4

ER -