Document Detail

Characterization of mini-protein S, a recombinant variant of protein S that lacks the sex hormone binding globulin-like domain.
MedLine Citation:
PMID:  9461535     Owner:  NLM     Status:  MEDLINE    
Protein S is a vitamin K-dependent glycoprotein involved in the regulation of the anticoagulant activity of activated protein C (APC). Also, an anticoagulant role for protein S, independent of APC, has been described. Protein S has a unique C-terminal sex hormone binding globulin (SHBG)-like domain that represents about half of the molecule. To define the role of this domain in APC cofactor activity and in binding to C4b-binding protein (C4BP), we have constructed a recombinant protein S molecule of N-terminal residues 1-242 that lacks the SHBG domain (mini-protein S). A panel of monoclonal antibodies directed against the N-terminal region of protein S recognized plasma-derived protein S, wild-type recombinant protein S and mini-protein S with similar affinities, whereas a monoclonal antibody that recognizes an epitope in the SHBG domain did not detect mini-protein S. Mini-protein S did not bind to C4BP in a solid-phase binding assay, and the cofactor activity of mini-protein S was not inhibited by preincubation with C4BP. In a plasma coagulation assay, the cofactor activity of mini-protein S was lower than wild-type or plasma-derived preparations. In contrast, no difference in APC cofactor activities was observed when the preparations were tested in purified systems that monitor the APC-mediated degradation of factors Va or VIIIa. In conclusion, we constructed a protein S molecule that fails to bind C4BP and still displays cofactor activity for APC. This confirms the role of the C-terminal SHBG region in C4BP binding and demonstrates that N-terminal residues 1-242 are sufficient for the expression of APC cofactor activity in a system using purified components. In plasma, however, the C-terminal SHBG region plays a role in the expression of optimal APC cofactor activity.
M Van Wijnen; J G Stam; G T Chang; J C Meijers; P H Reitsma; R M Bertina; B N Bouma
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Biochemical journal     Volume:  330 ( Pt 1)     ISSN:  0264-6021     ISO Abbreviation:  Biochem. J.     Publication Date:  1998 Feb 
Date Detail:
Created Date:  1998-04-09     Completed Date:  1998-04-09     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  389-96     Citation Subset:  IM    
Department of Haematology, G.03.647, University Hospital Utrecht, P. O. Box 85500, 3508 GA Utrecht, The Netherlands.
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MeSH Terms
Antibodies, Monoclonal
Blood Coagulation
Carrier Proteins / chemistry*
Cell Line
Factor VIIIa / metabolism
Factor Va / metabolism
Integrin alphaXbeta2
Protein Binding
Protein C / metabolism
Protein S / chemistry*
Recombinant Proteins
Sequence Deletion
Sex Hormone-Binding Globulin / chemistry*
Structure-Activity Relationship
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Carrier Proteins; 0/Integrin alphaXbeta2; 0/Protein C; 0/Protein S; 0/Recombinant Proteins; 0/Sex Hormone-Binding Globulin; 65522-14-7/Factor Va; 72175-66-7/Factor VIIIa

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