Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1983 Mar 1;209(3):857–863. doi: 10.1042/bj2090857

Degradation of human complement component C4b in the presence of the C4b-binding protein-protein S complex.

B Dahlbäck, B Hildebrand
PMCID: PMC1154166  PMID: 6223626

Abstract

Vitamin K-dependent protein S and the higher-molecular-weight form of C4b-binding protein (C4bp-high) interact, forming a 1:1 complex with a KD of approx. 1 X 10(-7) M [Dahlbäck (1983) Biochem. J. 209, 847-856]. In the present study the effect of protein S on the degradation of C4b by Factor I (C3b inactivator) and C4bp was investigated both in fluid phase and on cell surfaces, with the use of highly purified components. Fluid-phase degradation of C4b was monitored on sodium dodecyl sulphate/polyacrylamide-slab-gel electrophoresis, and the effect on surface-bound C4b was estimated by haemolytic assay. No effect of protein S could be demonstrated in any of the systems used. Thus, although bound to C4bp, protein S is neither involved in, nor does it affect, the interaction between C4bp and C4b. This indicates that the binding sites on the C4bp molecule for protein S and for C4b are independent and different.

Full text

PDF
857

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bolotin C., Morris S., Tack B., Prahl J. Purification and structural analysis of the fourth component of human complement. Biochemistry. 1977 May 3;16(9):2008–2015. doi: 10.1021/bi00628a039. [DOI] [PubMed] [Google Scholar]
  2. Campbell R. D., Dodds A. W., Porter R. R. The binding of human complement component C4 to antibody-antigen aggregates. Biochem J. 1980 Jul 1;189(1):67–80. doi: 10.1042/bj1890067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cooper N. R. Isolation and analysis of the mechanism of action of an inactivator of C4b in normal human serum. J Exp Med. 1975 Apr 1;141(4):890–903. [PMC free article] [PubMed] [Google Scholar]
  4. Crossley L. G., Porter R. R. Purification of the human complement control protein C3b inactivator. Biochem J. 1980 Oct 1;191(1):173–182. doi: 10.1042/bj1910173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  6. Dahlbäck B. Purification of human C4b-binding protein and formation of its complex with vitamin K-dependent protein S. Biochem J. 1983 Mar 1;209(3):847–856. doi: 10.1042/bj2090847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dahlbäck B. Purification of human vitamin K-dependent protein S and its limited proteolysis by thrombin. Biochem J. 1983 Mar 1;209(3):837–846. doi: 10.1042/bj2090837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dahlbäck B., Stenflo J. High molecular weight complex in human plasma between vitamin K-dependent protein S and complement component C4b-binding protein. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2512–2516. doi: 10.1073/pnas.78.4.2512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DiScipio R. G., Davie E. W. Characterization of protein S, a gamma-carboxyglutamic acid containing protein from bovine and human plasma. Biochemistry. 1979 Mar 6;18(5):899–904. doi: 10.1021/bi00572a026. [DOI] [PubMed] [Google Scholar]
  10. Fujita T., Gigli I., Nussenzweig V. Human C4-binding protein. II. Role in proteolysis of C4b by C3b-inactivator. J Exp Med. 1978 Oct 1;148(4):1044–1051. doi: 10.1084/jem.148.4.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fujita T., Nussenzweig V. The role of C4-binding protein and beta 1H in proteolysis of C4b and C3b. J Exp Med. 1979 Aug 1;150(2):267–276. doi: 10.1084/jem.150.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gigli I., Fujita T., Nussenzweig V. Modulation of the classical pathway C3 convertase by plasma proteins C4 binding protein and C3b inactivator. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6596–6600. doi: 10.1073/pnas.76.12.6596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gigli I., Porter R. R., Sim R. B. The unactivated form of the first component of human complement, C1. Biochem J. 1976 Sep 1;157(3):541–548. doi: 10.1042/bj1570541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harrison R. A., Thomas M. L., Tack B. F. Sequence determination of the thiolester site of the fourth component of human complement. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7388–7392. doi: 10.1073/pnas.78.12.7388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Iida K., Nussenzweig V. Complement receptor is an inhibitor of the complement cascade. J Exp Med. 1981 May 1;153(5):1138–1150. doi: 10.1084/jem.153.5.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Janatova J., Tack B. F. Fourth component of human complement: studies of an amine-sensitive site comprised of a thiol component. Biochemistry. 1981 Apr 28;20(9):2394–2402. doi: 10.1021/bi00512a005. [DOI] [PubMed] [Google Scholar]
  17. Kerr M. A. Limited proteolysis of complement components C2 and factor B. Structural analogy and limited sequence homology. Biochem J. 1979 Dec 1;183(3):615–622. doi: 10.1042/bj1830615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kisiel W. Human plasma protein C: isolation, characterization, and mechanism of activation by alpha-thrombin. J Clin Invest. 1979 Sep;64(3):761–769. doi: 10.1172/JCI109521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Law S. K., Lichtenberg N. A., Holcombe F. H., Levine R. P. Interaction between the labile binding sites of the fourth (C4) and fifth (C5) human complement proteins and erythrocyte cell membranes. J Immunol. 1980 Aug;125(2):634–639. [PubMed] [Google Scholar]
  20. Nagasawa S., Ichihara C., Stroud R. M. Cleavage of C4b by C3b inactivator: production of a nicked form of C4b, C4b', as an intermediate cleavage product of C4b by C3b inactivator. J Immunol. 1980 Aug;125(2):578–582. [PubMed] [Google Scholar]
  21. Nagasawa S., Stroud R. M. Cleavage of C2 by C1s into the antigenically distinct fragments C2a and C2b: demonstration of binding of C2b to C4b. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2998–3001. doi: 10.1073/pnas.74.7.2998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pangburn M. K., Schreiber R. D., Müller-Eberhard H. J. Human complement C3b inactivator: isolation, characterization, and demonstration of an absolute requirement for the serum protein beta1H for cleavage of C3b and C4b in solution. J Exp Med. 1977 Jul 1;146(1):257–270. doi: 10.1084/jem.146.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Porter R. R., Reid K. B. Activation of the complement system by antibody-antigen complexes: the classical pathway. Adv Protein Chem. 1979;33:1–71. doi: 10.1016/s0065-3233(08)60458-1. [DOI] [PubMed] [Google Scholar]
  24. Press E. M., Gagnon J. Human complement component C4. Structural studies on the fragments derived from C4b by cleavage with C3b inactivator. Biochem J. 1981 Nov 1;199(2):351–357. doi: 10.1042/bj1990351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Press E. M., Gagnon J. Human complement component C4. Structural studies on the fragments derived from C4b by cleavage with C3b inactivator. Biochem J. 1981 Nov 1;199(2):351–357. doi: 10.1042/bj1990351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Reid K. B., Porter R. R. The proteolytic activation systems of complement. Annu Rev Biochem. 1981;50:433–464. doi: 10.1146/annurev.bi.50.070181.002245. [DOI] [PubMed] [Google Scholar]
  27. Scharfstein J., Ferreira A., Gigli I., Nussenzweig V. Human C4-binding protein. I. Isolation and characterization. J Exp Med. 1978 Jul 1;148(1):207–222. doi: 10.1084/jem.148.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schreiber R. D., Müller-Eberhard H. J. Fourth component of human complement: description of a three polypeptide chain structure. J Exp Med. 1974 Nov 1;140(5):1324–1335. doi: 10.1084/jem.140.5.1324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shiraishi S., Stroud R. M. Cleavage products of C4b produced by enzymes in human serum. Immunochemistry. 1975 Dec;12(12):935–939. doi: 10.1016/0019-2791(75)90256-6. [DOI] [PubMed] [Google Scholar]
  30. Thorell J. I., Johansson B. G. Enzymatic iodination of polypeptides with 125I to high specific activity. Biochim Biophys Acta. 1971 Dec 28;251(3):363–369. doi: 10.1016/0005-2795(71)90123-1. [DOI] [PubMed] [Google Scholar]
  31. Villiers M. B., Reboul A., Thielens N. M., Colomb M. G. Purification and characterization of C4-binding protein from human serum. FEBS Lett. 1981 Sep 14;132(1):49–54. doi: 10.1016/0014-5793(81)80425-5. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES