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. 1968 Sep 1;128(3):533–551. doi: 10.1084/jem.128.3.533

THE SECOND COMPONENT OF HUMAN COMPLEMENT: ITS ISOLATION, FRAGMENTATION BY C'1 ESTERASE, AND INCORPORATION INTO C'3 CONVERTASE

Margaret J Polley 1, Hans J Müller-Eberhard 1
PMCID: PMC2138531  PMID: 5666963

Abstract

A method has been described for the purification and isolation of the second component of complement (C'2) from human serum. The protein is a β1-globulin with an approximate molecular weight of 117,000. Immunochemical analysis using a variety of specific antisera, including a monospecific antiserum to the isolated protein, indicate that the C'2 protein represents a heretofore unrecognized human serum constituent. Isolated C'2 contained 2 x 109 "effective molecules" per microgram and 1000 hemolytically active C'2 molecules were required to produce a single hemolytically effective C'2 site on erythrocytes undergoing immune cytolysis. C'1 esterase treatment of C'2 resulted in reduction of both its electrophoretic mobility and its molecular size, the latter observation indicating fragmentation of the molecule. Direct evidence was presented for the physical presence of C'2 as an integral part of the enzyme C'3 convertase.

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Selected References

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

  1. Cooper N. R., Müller-Eberhard H. J. A comparison of methods for the molecular quantitation of the fourth component of human complement. Immunochemistry. 1968 Mar;5(2):155–169. doi: 10.1016/0019-2791(68)90100-6. [DOI] [PubMed] [Google Scholar]
  2. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  3. Goudie R. B., Horne C. H., Wilkinson P. C. A simple method for producing antibody specific to a single selected diffusible antigen. Lancet. 1966 Dec 3;2(7475):1224–1226. doi: 10.1016/s0140-6736(66)92305-1. [DOI] [PubMed] [Google Scholar]
  4. HAINES A. L., LEPOW I. H. STUDIES ON HUMAN C'1-ESTERASE. I. PURIFICATION AND ENZYMATIC PROPERTIES. J Immunol. 1964 Mar;92:456–467. [PubMed] [Google Scholar]
  5. HAINES A. L., LEPOW I. H. STUDIES ON HUMAN C'1-ESTERASE. II. FUNCTION OF PURIFIED C'1-ESTERASE IN THE HUMAN COMPLEMENT SYSTEM. J Immunol. 1964 Mar;92:468–478. [PubMed] [Google Scholar]
  6. HJERTEN S., LEVIN O., TISELIUS A. Protein chromatography on calcium phosphate columns. Arch Biochem Biophys. 1956 Nov;65(1):132–155. doi: 10.1016/0003-9861(56)90183-7. [DOI] [PubMed] [Google Scholar]
  7. LEVINE L. Inhibition of immune hemolysis by diisopropyl fluorophosphate. Biochim Biophys Acta. 1955 Oct;18(2):283–284. doi: 10.1016/0006-3002(55)90069-x. [DOI] [PubMed] [Google Scholar]
  8. MUELLER-EBERHARD H. J., LEPOW I. H. C'1 ESTERASE EFFECT ON ACTIVITY AND PHYSICOCHEMICAL PROPERTIES OF THE FOURTH COMPONENT OF COMPLEMENT. J Exp Med. 1965 May 1;121:819–833. doi: 10.1084/jem.121.5.819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. NILSSON U. R., MUELLER-EBERHARD H. J. ISOLATION OF BETA IF-GLOBULIN FROM HUMAN SERUM AND ITS CHARACTERIZATION AS THE FIFTH COMPONENT OF COMPLEMENT. J Exp Med. 1965 Aug 1;122:277–298. doi: 10.1084/jem.122.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nelson R. A., Jr, Jensen J., Gigli I., Tamura N. Methods for the separation, purification and measurement of nine components of hemolytic complement in guinea-pig serum. Immunochemistry. 1966 Mar;3(2):111–135. doi: 10.1016/0019-2791(66)90292-8. [DOI] [PubMed] [Google Scholar]
  13. Polley M. J., Müller-Eberhard H. J. Enharncement of the hemolytic activity of the second component of human complement by oxidation. J Exp Med. 1967 Dec 1;126(6):1013–1025. doi: 10.1084/jem.126.6.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sitomer G., Stroud R. M., Mayer M. M. Reversible adsorption of C'2 by EAC'4: role of Mg2+, enumeration of competent SAC'4, two-step nature of C'2a fixation and estimation of its efficiency. Immunochemistry. 1966 Jan;3(1):57–69. doi: 10.1016/0019-2791(66)90282-5. [DOI] [PubMed] [Google Scholar]
  15. Stroud R. M., Mayer M. M., Miller J. A., McKenzie A. T. C'2ad, an inactive derivative of C'2 released during decay of EAC'4,2a. Immunochemistry. 1966 May;3(3):163–176. doi: 10.1016/0019-2791(66)90182-0. [DOI] [PubMed] [Google Scholar]

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