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Reevaluation of total, free, and bound protein-S and C4b-binding protein-levels in plasma anticoagulated with citrate or hirudin

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Overview

authors

  • Griffin, John
  • Gruber, A.
  • Fernandez, J. A.

publication date

  • June 1992

journal

  • Blood  Journal

abstract

  • 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.

subject areas

  • Anticoagulants
  • Carrier Proteins
  • Citrates
  • Citric Acid
  • Complement Inactivator Proteins
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Glycoproteins
  • Hirudins
  • Humans
  • Immunosorbent Techniques
  • Male
  • Molecular Weight
  • Partial Thromboplastin Time
  • Protein Binding
  • Protein S
  • Reference Values
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Identity

International Standard Serial Number (ISSN)

  • 0006-4971

PubMed ID

  • 1534488
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Additional Document Info

start page

  • 3203

end page

  • 3211

volume

  • 79

issue

  • 12

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