Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 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.

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

Selected References

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

  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]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES