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Reevaluation of total, free, and bound protein S and C4b-binding protein
levels in plasma anticoagulated with citrate or hirudin
JH Griffin, A Gruber and JA Fernandez
Department of Molecular and Experimental Medicine, Scripps Research
Institute, La Jolla, CA 92037.
The levels of total, free, and bound protein S and C4BP were determined
using enzyme-linked immunosorbent assays (ELISAs) in plasma samples (8
males and 8 females) that were individually subjected to immunoadsorption
studies in which "free protein S" was defined as that not adsorbed by
anti-C4BP antibody-Sepharose column and "free C4BP" as that not adsorbed by
anti-protein S antibody-Sepharose. Bound species were calculated as the
difference between total and free species. Free protein S (131 nmol/L)
averaged 38% of total protein S (346 nmol/L) and free C4BP (37 nmol/L)
averaged 14% of total C4BP (264 nmol/L) in plasma. There was an excellent
correlation between bound protein S and bound C4BP with 1:1 molar
stoichiometry and a good correlation between free protein S antigen and
protein S anticoagulant activity. It appears that free protein S is a
necessary consequence of the molar excess of protein S over C4BP. C4BP beta
chain antigen levels, measured using a new ELISA, averaged 218 nmol/L, a
value indistinguishable from the molar concentrations of bound protein S
(215 nmol/L) and bound C4BP (227 nmol/L). The free C4BP beta chain antigen
was only 4 nmol/L compared with 131 nmol/L free protein S. These results
suggest that free C4BP in plasma predominantly lacks the beta chain, that
almost all C4BP capable of binding protein S is associated with protein S,
and that the plasma levels of oligomeric forms of C4BP containing a beta
chain (alpha 7 beta and alpha 6 beta) that bind protein S directly and
stoichiometrically regulate free protein S levels.
Volume 79,
Issue 12,
pp. 3203-3211,
06/15/1992
Copyright © 1992 by The American Society of Hematology
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