HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Schwalbe, R. A. Articles by Nelsestuen, G. L. Search for Related Content PubMed PubMed Citation Articles by Schwalbe, R. A. Articles by Nelsestuen, G. L. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 264, Issue 34, 20288-20296, 12, 1989

Protein structural requirements and properties of membrane binding by gamma-carboxyglutamic acid-containing plasma proteins and peptides

RA Schwalbe, J Ryan, DM Stern, W Kisiel, B Dahlback and GL Nelsestuen
Department of Biochemistry, University of Minnesota, St. Paul 55108.

The membrane-binding characteristics of a number of modified vitamin K- dependent proteins and peptides showed a general pattern of structural requirements. The amino-terminal peptides from human prothrombin (residues 1-41 and 1-44, 60:40) bovine factor X (residues 1-44), and bovine factor IX (residues 1-42), showed a general requirement for a free amino-terminal group, an intact disulfide, and the tyrosine homologous to Tyr44 of factor X for membrane binding. Consequently, the peptide from factor IX did not bind to membranes. Any of several modifications of the amino terminus, except reaction with trinitrobenzenesulfonic acid, abolished membrane binding by the factor X and prothrombin peptides. Calcium, but not magnesium, protected the amino terminus from chemical modification. The requirement for a free amino terminus was also shown to be true for intact prothrombin fragment 1, factor X, and factor IX. Although aggregation of the peptide-vesicle complexes greatly complicated accurate estimation of equilibrium binding constants, results with the factor X peptide indicated an affinity that was not greatly different from that of the parent protein. The most striking difference shown by the peptides was a requirement for about 10 times as much calcium as the parent proteins. In a manner similar to the parent proteins, the prothrombin and factor X peptides showed a large calcium-dependent quenching of tryptophan fluorescence. This fluorescence quenching in the peptides also required about 10 times the calcium needed by the parent proteins. Thus, the 1-45 region of the vitamin K-dependent proteins contained most of the membrane-binding structure but lacked component(s) needed for high affinity calcium binding. Protein S that was modified by thrombin cleavage at Arg52 and Arg70 showed approximately the same behavior as the amino-terminal 45-residue peptides. That is, it bound to membranes with overall affinity that was similar to native protein S but required high calcium concentrations. These results suggested that the second disulfide loop of protein S (Cys47-Cys72) and prothrombin (Cys48-Cys61) were involved in high affinity calcium binding. Since factor X lacks a homologous disulfide loop, an alternative structure must serve a similar function. A striking property of protein S was dissociation from membranes by high calcium. While this property was shared by all the vitamin K-dependent proteins, protein S showed this most dramatically and supported protein-membrane binding by calcium bridging.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				L. Kask, L. A. Trouw, B. Dahlback, and A. M. Blom The C4b-binding Protein-Protein S Complex Inhibits the Phagocytosis of Apoptotic Cells J. Biol. Chem., June 4, 2004; 279(23): 23869 - 23873. [Abstract] [Full Text] [PDF]

				H. Mizuno, Z. Fujimoto, H. Atoda, and T. Morita Crystal structure of an anticoagulant protein in complex with the Gla domain of factor X PNAS, June 7, 2001; (2001) 131179698. [Abstract] [Full Text] [PDF]

				H. Isawa, M. Yuda, K. Yoneda, and Y. Chinzei The Insect Salivary Protein, Prolixin-S, Inhibits Factor IXa Generation and Xase Complex Formation in the Blood Coagulation Pathway J. Biol. Chem., February 25, 2000; 275(9): 6636 - 6641. [Abstract] [Full Text] [PDF]

				S. J. Freedman, M. D. Blostein, J. D. Baleja, M. Jacobs, B. C. Furie, and B. Furie Identification of the Phospholipid Binding Site in the Vitamin K-dependent Blood Coagulation Protein Factor IX J. Biol. Chem., July 5, 1996; 271(27): 16227 - 16236. [Abstract] [Full Text] [PDF]

				L. M. O'Brien, L. V. Medved, and P. J. Fay Localization of Factor IXa and Factor VIIIa Interactive Sites J. Biol. Chem., November 10, 1995; 270(45): 27087 - 27092. [Abstract] [Full Text] [PDF]

				S. J. Freedman, B. C. Furie, B. Furie, and J. D. Baleja Structure of the Metal-free [IMAGE]-Carboxyglutamic Acid-rich Membrane Binding Region of Factor IX by Two-dimensional NMR Spectroscopy J. Biol. Chem., April 7, 1995; 270(14): 7980 - 7987. [Abstract] [Full Text] [PDF]

				M. D. Blostein, A. C. Rigby, M. Jacobs, B. Furie, and B. C. Furie The Gla Domain of Human Prothrombin Has a Binding Site for Factor Va J. Biol. Chem., November 22, 2000; 275(48): 38120 - 38126. [Abstract] [Full Text] [PDF]

				L. A. Falls, B. C. Furie, M. Jacobs, B. Furie, and A. C. Rigby The omega -Loop Region of the Human Prothrombin gamma -Carboxyglutamic Acid Domain Penetrates Anionic Phospholipid Membranes J. Biol. Chem., June 22, 2001; 276(26): 23895 - 23902. [Abstract] [Full Text] [PDF]

				A. Srivastava, J. Wang, R. Majumder, A. R. Rezaie, J. Stenflo, C. T. Esmon, and B. R. Lentz Localization of Phosphatidylserine Binding Sites to Structural Domains of Factor Xa J. Biol. Chem., January 11, 2002; 277(3): 1855 - 1863. [Abstract] [Full Text] [PDF]

				H. Mizuno, Z. Fujimoto, H. Atoda, and T. Morita Crystal structure of an anticoagulant protein in complex with the Gla domain of factor X PNAS, June 19, 2001; 98(13): 7230 - 7234. [Abstract] [Full Text] [PDF]