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Binding site for C4b-binding protein in vitamin K-dependent protein S fully contained in carboxy-terminal laminin-G-type repeats. A study using recombinant factor IX-protein S chimeras and surface plasmon resonance.

Publication ,  Journal Article
He, X; Shen, L; Malmborg, AC; Smith, KJ; Dahlback, B; Linse, S
Published in: Biochemistry
March 25, 1997

The interaction between vitamin K-dependent protein S and the C4b-binding protein (C4BP) was studied using surface plasmon resonance and genetic engineering. The affinity, as well as association and dissociation rates of the complex, was measured for human and bovine protein S at five different calcium concentrations. The binding to C4BP of six protein hybrids containing different parts of coagulation factor IX and protein S was studied in the absence and presence of calcium. The results show that dissociation of the human protein S-C4BP complex is extremely slow in the presence of > or = 10 microM calcium (k(off) = 7 x 10(-6) s(-1)) and the association rate constant is k(on) = 7 x 10(4) M(-1) s(-1). Human and bovine protein S were found to bind to human C4BP with the same affinity, K(D) = 0.1 nM, but the rates of association and dissociation were higher for the bovine protein S (k(on) = 2 x 10(5) M(-1) s(-1), k(off) = 2 x 10(-5) s(-1)). In the absence of calcium, the affinity for C4BP was reduced by a factor of 65 for human protein S and by a factor of 40 for bovine protein S. The decreased affinity could be mainly attributed to an increased off-rate (12-17-fold), while the on-rate decreased 3-4-fold. The studies using chimeric proteins show that the portion of protein S that is responsible for binding to C4BP is fully contained in the two laminin-G-type repeats, which are homologous to the sex hormone binding globulin (SHBG). All hybrids that contain the laminin-G-type repeats bind to C4BP with the same affinity as recombinant protein S, whereas hybrids lacking these repeats show no detectable binding to C4BP. The present data also suggest that the effect of calcium on the C4BP-binding properties is mediated by calcium binding site(s) in the laminin-G-type repeats.

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Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

March 25, 1997

Volume

36

Issue

12

Start / End Page

3745 / 3754

Location

United States

Related Subject Headings

  • Recombinant Proteins
  • Recombinant Fusion Proteins
  • Receptors, Laminin
  • Receptors, Complement
  • Protein S
  • Protein Precursors
  • Protein Conformation
  • Polymerase Chain Reaction
  • Molecular Sequence Data
  • Models, Molecular
 

Citation

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He, X., Shen, L., Malmborg, A. C., Smith, K. J., Dahlback, B., & Linse, S. (1997). Binding site for C4b-binding protein in vitamin K-dependent protein S fully contained in carboxy-terminal laminin-G-type repeats. A study using recombinant factor IX-protein S chimeras and surface plasmon resonance. Biochemistry, 36(12), 3745–3754. https://doi.org/10.1021/bi962315q
He, X., L. Shen, A. C. Malmborg, K. J. Smith, B. Dahlback, and S. Linse. “Binding site for C4b-binding protein in vitamin K-dependent protein S fully contained in carboxy-terminal laminin-G-type repeats. A study using recombinant factor IX-protein S chimeras and surface plasmon resonance.Biochemistry 36, no. 12 (March 25, 1997): 3745–54. https://doi.org/10.1021/bi962315q.
Journal cover image

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

March 25, 1997

Volume

36

Issue

12

Start / End Page

3745 / 3754

Location

United States

Related Subject Headings

  • Recombinant Proteins
  • Recombinant Fusion Proteins
  • Receptors, Laminin
  • Receptors, Complement
  • Protein S
  • Protein Precursors
  • Protein Conformation
  • Polymerase Chain Reaction
  • Molecular Sequence Data
  • Models, Molecular