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. 1981 Dec 1;199(3):485–496. doi: 10.1042/bj1990485

The binding of human complement proteins C5, factor B, beta 1H and properdin to complement fragment C3b on zymosan.

R G DiScipio
PMCID: PMC1163402  PMID: 6462133

Abstract

The covalent binding of complement fragment C3b to zymosan by the action of the alternative-pathway C3 convertase and the reversible binding of several complement proteins (component C5, factor B, beta 1H and properdin) to C3b on zymosan have been investigated. When C3b is deposited on zymosan after activation by a surface-bound C3 convertase, the C3b molecules are deposited in foci around the C3 convertase site, with an average of 30 C3b molecules per site. The association constants of C5, factor B, beta 1H, and properdin for C3b bound to zymosan have been determined. The association constants ranged from 6.5 x 10(-5) M-1 for factor B to 2.9 x 10(7) M-1 for properdin. An approximate stoichiometry of 1 : 1 for C5, factor B, and properdin binding to C3b has been observed. Curvilinear Scatchard plots were observed for beta 1H binding to C3b, with the maximal extrapolated ratio of beta 1H to C3b of 0.32. Physiological amounts of properdin increase by 7-fold the affinity constant for factor B binding to C3b with no alteration in the stoichiometry. Similarly, physiological amounts of factor B increase the affinity constant of properdin to C3b about 4-fold with only a small measured difference in stoichiometry. Competition binding studies and protein modification suggest that C5, factor B, beta 1H, and properdin each bind to a distinct region on C3b.

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Selected References

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  1. Bianco C., Patrick R., Nussenzweig V. A population of lymphocytes bearing a membrane receptor for antigen-antibody-complement complexes. I. Separation and characterization. J Exp Med. 1970 Oct 1;132(4):702–720. doi: 10.1084/jem.132.4.702. [DOI] [PMC free article] [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. Conrad D. H., Carlo J. R., Ruddy S. Interaction of beta1H globulin with cell-bound C3b: quantitative analysis of binding and influence of alternative pathway components on binding. J Exp Med. 1978 Jun 1;147(6):1792–1805. doi: 10.1084/jem.147.6.1792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cooper N. R., Müller-Eberhard H. J. The reaction mechanism of human C5 in immune hemolysis. J Exp Med. 1970 Oct 1;132(4):775–793. doi: 10.1084/jem.132.4.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Curman B., Sandberg-Trägårdh L., Peterson P. A. Chemical characterization of human factor B of the alternate pathway of complement activation. Biochemistry. 1977 Nov 29;16(24):5368–5375. doi: 10.1021/bi00643a031. [DOI] [PubMed] [Google Scholar]
  8. David G. S. Solid state lactoperoxidase: a highly stable enzyme for simple, gentle iodination of proteins. Biochem Biophys Res Commun. 1972 Jul 25;48(2):464–471. doi: 10.1016/s0006-291x(72)80074-3. [DOI] [PubMed] [Google Scholar]
  9. Di Scipio R. G., Hermodson M. A., Yates S. G., Davie E. W. A comparison of human prothrombin, factor IX (Christmas factor), factor X (Stuart factor), and protein S. Biochemistry. 1977 Feb 22;16(4):698–706. doi: 10.1021/bi00623a022. [DOI] [PubMed] [Google Scholar]
  10. Dierich M. P., Bokisch V. A. Receptor-binding sites on C3 and C3b. J Immunol. 1977 Jun;118(6):2145–2150. [PubMed] [Google Scholar]
  11. 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]
  12. Götze O., Medicus R. G., Müller-Eberhard H. J. Alternative pathway of complement: nonenzymatic, reversible transition of precursor to active properdin. J Immunol. 1977 Feb;118(2):525–532. [PubMed] [Google Scholar]
  13. Harpel P. C., Hayes M. B., Hugli T. E. Heat-induced fragmentation of human alpha 2-macroglobulin. J Biol Chem. 1979 Sep 10;254(17):8669–8678. [PubMed] [Google Scholar]
  14. Howard J. B. Methylamine reaction and denaturation-dependent fragmentation of complement component 3. Comparison with alpha2-macroglobulin. J Biol Chem. 1980 Aug 10;255(15):7082–7084. [PubMed] [Google Scholar]
  15. Howard J. B., Vermeulen M., Swenson R. P. The temperature-sensitive bond in human alpha 2-macroglobulin is the alkylamine-reactive site. J Biol Chem. 1980 May 10;255(9):3820–3823. [PubMed] [Google Scholar]
  16. Isenman D. E., Podack E. R., Cooper N. R. The interaction of C5 with C3b in free solution: a sufficient condition for cleavage by a fluid phase C3/C5 convertase. J Immunol. 1980 Jan;124(1):326–331. [PubMed] [Google Scholar]
  17. Isenman D. E., Sundsmo J. S., Cooper N. R. The effects of mild reduction on the structure and function of C3. J Immunol. 1980 Oct;125(4):1798–1805. [PubMed] [Google Scholar]
  18. Janatova J., Lorenz P. E., Schechter A. N., Prahl J. W., Tack B. F. Third component of human complement: appearance of a sulfhydryl group following chemical or enzymatic inactivation. Biochemistry. 1980 Sep 16;19(19):4471–4478. [PubMed] [Google Scholar]
  19. Kazatchkine M. D., Fearon D. T., Austen K. F. Human alternative complement pathway: membrane-associated sialic acid regulates the competition between B and beta1 H for cell-bound C3b. J Immunol. 1979 Jan;122(1):75–81. [PubMed] [Google Scholar]
  20. Law S. K., Fearon D. T., Levine R. P. Action of the C3b-inactivator on the cell-bound C3b. J Immunol. 1979 Mar;122(3):759–765. [PubMed] [Google Scholar]
  21. Law S. K., Levine R. P. Interaction between the third complement protein and cell surface macromolecules. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2701–2705. doi: 10.1073/pnas.74.7.2701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Law S. K., Lichtenberg N. A., Levine R. P. Evidence for an ester linkage between the labile binding site of C3b and receptive surfaces. J Immunol. 1979 Sep;123(3):1388–1394. [PubMed] [Google Scholar]
  24. Lay W. H., Nussenzweig V. Receptors for complement of leukocytes. J Exp Med. 1968 Nov 1;128(5):991–1009. doi: 10.1084/jem.128.5.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mardiney M. R., Jr, Müller-Eberhard H. J., Feldman J. D. Ultrastructural localization of the third and fourth components of complement on complement-cell complexes. Am J Pathol. 1968 Aug;53(2):253–261. [PMC free article] [PubMed] [Google Scholar]
  26. Medicus R. G., Esser A. F., Fernandez H. N., Müller-Eberhard H. J. Native and activated properdin: interconvertibility and identity of amino- and carboxy-terminal sequences. J Immunol. 1980 Feb;124(2):602–606. [PubMed] [Google Scholar]
  27. Medicus R. G., Götze O., Müller-Eberhard H. J. Alternative pathway of complement: recruitment of precursor properdin by the labile C3/C5 convertase and the potentiation of the pathway. J Exp Med. 1976 Oct 1;144(4):1076–1093. doi: 10.1084/jem.144.4.1076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Minta J. O., Lepow I. H. Studies on the sub-unit structure of human properdin. Immunochemistry. 1974 Jul;11(7):361–368. doi: 10.1016/0019-2791(74)90189-x. [DOI] [PubMed] [Google Scholar]
  29. Molenaar J. L., Müller M. A., Engelfriet C. P., Pondman K. W. Changes in antigenic properties of human C3 upon activation and conversion by trypsin. J Immunol. 1974 Apr;112(4):1444–1451. [PubMed] [Google Scholar]
  30. Müller-Eberhard H. J., Götze O. C3 proactivator convertase and its mode of action. J Exp Med. 1972 Apr 1;135(4):1003–1008. doi: 10.1084/jem.135.4.1003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Müller-Eberhard H. J., Schreiber R. D. Molecular biology and chemistry of the alternative pathway of complement. Adv Immunol. 1980;29:1–53. doi: 10.1016/s0065-2776(08)60042-5. [DOI] [PubMed] [Google Scholar]
  32. Nagaki K., Iida K., Okubo M., Inai S. Reaction mechanisms of beta1H globulin. Int Arch Allergy Appl Immunol. 1978;57(3):221–232. doi: 10.1159/000232106. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Nagasawa S., Stroud R. M. Mechanism of action of the C3b inactivator: requirement for a high molecular weight cofactor (C3b-C4bINA cofactor) and production of a new C3b derivative (C3b'). Immunochemistry. 1977 Nov-Dec;14(11-12):749–756. doi: 10.1016/0019-2791(77)90345-7. [DOI] [PubMed] [Google Scholar]
  35. Pangburn M. K., Müller-Eberhard H. J. Complement C3 convertase: cell surface restriction of beta1H control and generation of restriction on neuraminidase-treated cells. Proc Natl Acad Sci U S A. 1978 May;75(5):2416–2420. doi: 10.1073/pnas.75.5.2416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Parkes C., DiScipio R. G., Kerr M. A., Prohaska R. The separation of functionally distinct forms of the third component of human complement (C3). Biochem J. 1981 Mar 1;193(3):963–970. doi: 10.1042/bj1930963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ross G. D., Polley M. J. Specificity of human lymphocyte complement receptors. J Exp Med. 1975 May 1;141(5):1163–1180. doi: 10.1084/jem.141.5.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Salvesen G. S., Barrett A. J. Covalent binding of proteinases in their reaction with alpha 2-macroglobulin. Biochem J. 1980 Jun 1;187(3):695–701. doi: 10.1042/bj1870695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sottrup-Jensen L., Petersen T. E., Magnusson S. A thiol-ester in alpha 2-macroglobulin cleaved during proteinase complex formation. FEBS Lett. 1980 Dec 1;121(2):275–279. doi: 10.1016/0014-5793(80)80361-9. [DOI] [PubMed] [Google Scholar]
  41. Swenson R. P., Howard J. B. Characterization of alkylamine-sensitive site in alpha 2-macroglobulin. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4313–4316. doi: 10.1073/pnas.76.9.4313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tack B. D., Prahl J. W. Third component of human complement: purification from plasma and physicochemical characterization. Biochemistry. 1976 Oct 5;15(20):4513–4521. doi: 10.1021/bi00665a028. [DOI] [PubMed] [Google Scholar]
  43. Tack B. F., Harrison R. A., Janatova J., Thomas M. L., Prahl J. W. Evidence for presence of an internal thiolester bond in third component of human complement. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5764–5768. doi: 10.1073/pnas.77.10.5764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tack B. F., Morris S. C., Prahl J. W. Fifth component of human complement: purification from plasma and polypeptide chain structure. Biochemistry. 1979 Apr 17;18(8):1490–1497. doi: 10.1021/bi00575a016. [DOI] [PubMed] [Google Scholar]
  45. Vogt W., Dames W., Schmidt G., Dieminger L. Complement activation by the properdin system: formation of a stoichiometric. C3 cleaving complex of properdin factor B with C36. Immunochemistry. 1977 Mar;14(3):201–205. doi: 10.1016/0019-2791(77)90195-1. [DOI] [PubMed] [Google Scholar]
  46. Vogt W., Schmidt G., Hinsch B. Interference of propamidine with binding of the fifth component of complement to surface-fixed C3b, and with C5 activation. Immunology. 1979 Jan;36(1):139–143. [PMC free article] [PubMed] [Google Scholar]
  47. Vogt W., Schmidt G., Von Buttlar B., Dieminger L. A new function of the activated third component of complement: binding to C5, an essential step for C5 activation. Immunology. 1978 Jan;34(1):29–40. [PMC free article] [PubMed] [Google Scholar]
  48. Whaley K., Ruddy S. Modulation of the alternative complement pathways by beta 1 H globulin. J Exp Med. 1976 Nov 2;144(5):1147–1163. doi: 10.1084/jem.144.5.1147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Zemel E. S., Cartwright-Harwick C. M., Nilsson U. R. Effect of reduction on the structural, biologic, and immunologic properties of the third component of human complement. J Immunol. 1980 Sep;125(3):1099–1103. [PubMed] [Google Scholar]

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