Document Detail


Expression and characterization of a recombinant C4b-binding protein lacking the beta-chain.
MedLine Citation:
PMID:  8948435     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
C4b-binding protein (C4BP) is a high-molecular-mass glycoprotein which contains binding sites for complement component C4b, anti-coagulant vitamin K-dependent protein S and serum amyloid P component (SAP). The major form of C4BP in plasma is composed of seven identical alpha-chains and a single beta-chain. We have expressed full-length cDNA for the alpha-chain in a eukaryotic expression system and characterized functional properties of non-beta-chain-containing C4BP. During synthesis, recombinant alpha-chains polymerized into two different high-molecular-mass C4BP forms which were composed of seven or eight alpha-chains. Recombinant C4BP bound C4(H2O) (used instead of C4b) equally as well as native C4BP, functioned equally as well as factor I cofactor in the degradation of C4(H2O) and bound to SAP. In contrast, the recombinant C4BP did not bind protein S and therefore did not inhibit the ability of protein S to function as a cofactor to activated protein C. Tunicamycin treatment of the transfected cells prevented N-linked glycosylation, but did not affect polymerization of the alpha-chains into a high-molecular-mass C4BP. The non-glycosylated C4BP had comparable properties to glycosylated C4BP in several functional assays. These results demonstrate polymerization of C4BP alpha-chains to be independent both of the beta-chain and of the N-linked carbohydrates. Moreover, N-linked carbohydrates and the beta-chain were neither required for the ability of C4BP to bind C4b and to function as factor I cofactor nor for the interaction with SAP.
Authors:
Y Härdig; P García de Frutos; B Dahlbäck
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Biochemical journal     Volume:  308 ( Pt 3)     ISSN:  0264-6021     ISO Abbreviation:  Biochem. J.     Publication Date:  1995 Jun 
Date Detail:
Created Date:  1997-01-07     Completed Date:  1997-01-07     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  795-800     Citation Subset:  IM    
Affiliation:
Department of Clinical Chemistry, University of Lund, Malmö General Hospital, Sweden.
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MeSH Terms
Descriptor/Qualifier:
Binding, Competitive
Blood Coagulation / drug effects,  physiology
Blood Proteins / chemistry,  metabolism
Chromatography, Affinity
Cloning, Molecular
Complement Inactivator Proteins*
DNA, Complementary / genetics
Electrophoresis, Polyacrylamide Gel
Fibrinogen / metabolism
Gene Expression Regulation
Glycoproteins*
Glycosylation
Humans
Protein Binding
Protein Conformation
Protein S / metabolism
Receptors, Complement / chemistry,  genetics,  metabolism*
Recombinant Proteins / genetics,  isolation & purification
Serum Amyloid P-Component / metabolism
Transfection / genetics
Tunicamycin / pharmacology
Chemical
Reg. No./Substance:
0/Blood Proteins; 0/Complement Inactivator Proteins; 0/DNA, Complementary; 0/Glycoproteins; 0/Protein S; 0/Receptors, Complement; 0/Recombinant Proteins; 0/Serum Amyloid P-Component; 11089-65-9/Tunicamycin; 9001-32-5/Fibrinogen
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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