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Thromb Haemost 1990; 64(02): 245-250
DOI: 10.1055/s-0038-1647294
Original Article
Schattauer GmbH Stuttgart

Identification of Different Forms of Human C4b-Binding Protein Lacking β-Chain and Protein S Binding Ability

Martin Hessing
The Department of Haematology, University Hospital Utrecht, Utrecht, The Netherlands
,
Deon Kanters
The Department of Haematology, University Hospital Utrecht, Utrecht, The Netherlands
,
Tilman M Hackeng
The Department of Haematology, University Hospital Utrecht, Utrecht, The Netherlands
,
Bonno N Bouma
The Department of Haematology, University Hospital Utrecht, Utrecht, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 01 March 1990

Accepted after revision04 May 1990

Publication Date:
25 July 2018 (online)

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Summary

Human C4b-binding protein (C4BP) is a multimeric regulatory component of the complement system that circulates in plasma either as a free protein or in a noncovalent complex with the vitamin K-dependent protein S. The major form of C4BP is composed of seven identical α-chains (70 kDa) and one β-chain (45 kDa). C4BP was purified from human plasma after barium citrate adsorption using anti-C4BP monoclonal antibody affinity chromatography. C4BP-high and low Mr forms were both obtained from the barium citrate precipitate and the supernatant. C4BP-high and low forms from the barium citrate precipitate were separated by sodium dodecylsulfate polyacrylamide slab gel electrophoresis and extracted with Triton X-100. Both forms contained the β-chain as was demonstrated on sodium dodecylsulfate polyacrylamide slab gel electrophoresis under reduced conditions after silverstaining and with Western-blotting using monoclonal antibodies specific for the β-chain. The C4BP-high and low forms demonstrated similar protein S binding affinity (KA: 3.18 × 108 and 3.21 × 108 M-1, respectively) in a C4BP-protein S binding assay and a protein S ligand blot using a peroxidase-conjugated monoclonal anti-protein S antibody. The barium citrate supernatant contained two forms of C4BP-high and one form of C4BP-low. One form of C4BP-high did contain the β-chain and was capable of protein S binding (KA: 4.35 × 108 M-1). The two other forms of C4BP lacked the β-chain and were unable to bind protein S.

 

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