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. 1977 Oct 1;146(4):1001–1018. doi: 10.1084/jem.146.4.1001

Purification and characterization of mouse serum protein with specific binding affinity for C4 (Ss protein)

A Ferreira, M Takahashi, V Nussenzweig
PMCID: PMC2180828  PMID: 894192

Abstract

A new component of the complement (C).system, with a specific binding affinity for the activated Ss-protein (C4) has been identified in mouse serum. This protein, named Ss- (or C4)-binding protein (Ss-bp), was purified about 200 times from mouse plasma. Ss-bp is a heat stable (56 degrees C, 60 rain) β-globulin with a sedimentation coefficient in sucrose density ultracentrifugation of 10s. Its concentration in serum of adult male and female mice is 160 and 60 μg/ml, respectively. In EDTA-plasma, Ss and Ss-bp are not associated and can be separated by chromatography in Sephadex G-200. However, in serum Ss-bp binds tightly to Ss. The bonds between these proteins cannot be reversed by chelation of divalent cations. As a consequence of the formation of Ss/Ss-bp complexes, the properties of Ss-bp appear to be quite different in serum of mice with high (Ss-H) or low (Ss-L) levels of Ss-protein. In Ss-H serum, all of Ss- bp is bound to Ss. In Ss-L serum, Ss-bp is mostly free. Because the electrophoretic mobilities of free and complexed Ss-bp are quite different, Ss-bp appears to be polymorphic in serum (but not in EDTA- plasma). The strict dependency of the apparent electrophoretic mobility of Ss-bp on the levels of Ss in serum was demonstrated in a series of congenic mice and among the progeny of a cross between Ss-H and Ss-L strains of mice. Without exception, the slow and fast varieties of Ss-bp were associated with the Ss-L and Ss-H traits. Ss-bp of the slow variety can be transformed into the fast variety by addition of pure human C4, or C4-sufficient guinea pig serum, to Ss-L serum. In both instances Ss-bp formed stable complexes with C4 or a C4- derived peptide. These findings highlight the binding specificity of Ss- bp for the fourth component of the complement system, and in addition they demonstrate a functional homology between the Ss-protein and C4 from two different species.

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Selected References

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  1. CINADER B., DUBISKI S., WARDLAW A. C. DISTRIBUTION, INHERITANCE, AND PROPERTIES OF AN ANTIGEN, MUB1, AND ITS RELATION TO HEMOLYTIC COMPLEMENT. J Exp Med. 1964 Nov 1;120:897–924. doi: 10.1084/jem.120.5.897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Caren L. D., Rosenberg L. T. Steroids and serum complement in mice: influence of hydrocortisone, diethylstilbestrol, and testosterone. Science. 1966 May 6;152(3723):782–783. doi: 10.1126/science.152.3723.782. [DOI] [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. Curman B., Ostberg L., Sandberg L., Malmheden-Eriksson I., Stålenheim G., Rask L., Peterson P. A. H-2 linked Ss protein is C4 component of complement. Nature. 1975 Nov 20;258(5532):243–245. doi: 10.1038/258243a0. [DOI] [PubMed] [Google Scholar]
  5. Eden A., Bianco C., Nussenzweig V. A population of lymphocytes bearing a membrane receptor antigen-antibody-complement complexes. II. Specific isolation. Cell Immunol. 1971 Dec;2(6):658–669. doi: 10.1016/0008-8749(71)90013-x. [DOI] [PubMed] [Google Scholar]
  6. Ferreira A., Nussenweig V. Control of C3 levels in mice during ontogeny by a gene in the central region of the H-2 complex. Nature. 1976 Apr 15;260(5552):613–615. doi: 10.1038/260613a0. [DOI] [PubMed] [Google Scholar]
  7. Gigli I., Ruddy S., Austen K. F. The stoichiometric measurement of the serum inhibition of the first component of complement by the inhibition of immune hemolysis. J Immunol. 1968 Jun;100(6):1154–1164. [PubMed] [Google Scholar]
  8. Goldman M. B., Goldman J. N. Relationship of functional levels of early components of complement to the H-2 complex of mice. J Immunol. 1976 Nov;117(5 Pt 1):1584–1588. [PubMed] [Google Scholar]
  9. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  10. Hansen T. H., Krasteff T. N., Shreffler D. C. Quantitative variations in the expression of the mouse serum antigen Ss and its sex-limited allotype Slp. Biochem Genet. 1974 Oct;12(4):281–293. doi: 10.1007/BF00485949. [DOI] [PubMed] [Google Scholar]
  11. Hansen T. H., Shin H. S., Shreffler D. C. Evidence for the involvement of the Ss protein of the mouse in the hemolytic complement system. J Exp Med. 1975 May 1;141(5):1216–1220. doi: 10.1084/jem.141.5.1216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hansen T. H., Shreffler D. C. Characterization of a constitutive variant of the murine serum protein allotype, Slp. J Immunol. 1976 Nov;117(5 Pt 1):1507–1513. [PubMed] [Google Scholar]
  13. Hinzová E., Démant P., Iványi P. Genetic control of haemolytic complement in mice: association with H-2. Folia Biol (Praha) 1972;18(4):237–243. [PubMed] [Google Scholar]
  14. LEVY L. R., LEPOW I. H. Assay and properties of serum inhibitor of C'l-esterase. Proc Soc Exp Biol Med. 1959 Aug-Sep;101:608–611. doi: 10.3181/00379727-101-25034. [DOI] [PubMed] [Google Scholar]
  15. Lachmann P. J., Grennan D., Martin A., Demant P. Identification of Ss protein as murine C4. Nature. 1975 Nov 20;258(5532):242–243. doi: 10.1038/258242a0. [DOI] [PubMed] [Google Scholar]
  16. MARDINEY M. R., Jr, MUELLER-EBERHARD H. J. MOUSE BETA-1C-GLOBULIN: PRODUCTION OF ANTISERUM AND CHARACTERIZATION IN THE COMPLEMENT REACTION. J Immunol. 1965 Jun;94:877–882. [PubMed] [Google Scholar]
  17. Mantovani B., Rabinovitch M., Nussenzweig V. Phagocytosis of immune complexes by macrophages. Different roles of the macrophage receptor sites for complement (C3) and for immunoglobulin (IgG). J Exp Med. 1972 Apr 1;135(4):780–792. doi: 10.1084/jem.135.4.780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Meo T., Krasteff T., Shreffler D. C. Immunochemical characterization of murine H-2 controlled Ss (serum substance) protein through identification of its human homologue as the fourth component of complement. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4536–4540. doi: 10.1073/pnas.72.11.4536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Müller-Eberhard H. J. Complement. Annu Rev Biochem. 1975;44:697–724. doi: 10.1146/annurev.bi.44.070175.003405. [DOI] [PubMed] [Google Scholar]
  20. Müller-Eberhard H. J., Polley M. J., Calcott M. A. Formation and functional significance of a molecular complex derived from the second and the fourth component of human complement. J Exp Med. 1967 Feb 1;125(2):359–380. doi: 10.1084/jem.125.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Passmore H. C., Shreffler D. C. A sex-limited serum protein variant in the mouse: hormonal control of phenotypic expression. Biochem Genet. 1971 Apr;5(2):201–209. doi: 10.1007/BF00485645. [DOI] [PubMed] [Google Scholar]
  22. Passmore H. C., Shreffler D. C. A sex-limited serum protein variant in the mouse: inheritance and association with the H-2 region. Biochem Genet. 1970 Jun;4(3):351–365. doi: 10.1007/BF00485752. [DOI] [PubMed] [Google Scholar]
  23. Ruddy S., Austen K. F. C3b inactivator of man. II. Fragments produced by C3b inactivator cleavage of cell-bound or fluid phase C3b. J Immunol. 1971 Sep;107(3):742–750. [PubMed] [Google Scholar]
  24. 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]
  25. Shreffler D C, Owen R D. A Serologically Detected Variant in Mouse Serum: Inheritance and Association with the Histocompatibility-2 Locus. Genetics. 1963 Jan;48(1):9–25. doi: 10.1093/genetics/48.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shreffler D. C., David C. S. Studies on recombination within the mouse H-2 complex. I. Three recombinants which position the Ss locus within the complex. Tissue Antigens. 1972;2(3):232–240. doi: 10.1111/j.1399-0039.1972.tb00140.x. [DOI] [PubMed] [Google Scholar]
  27. Weiler J. M., Daha M. R., Austen K. F., Fearon D. T. Control of the amplification convertase of complement by the plasma protein beta1H. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3268–3272. doi: 10.1073/pnas.73.9.3268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Weintraub R. M., Churchill W. H., Jr, Crisler C., Rapp H. J., Borsos T. Mouse complement: influence of sex hormones on its activity. Science. 1966 May 6;152(3723):783–785. doi: 10.1126/science.152.3723.783. [DOI] [PubMed] [Google Scholar]
  29. 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]

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