Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1975 Jun 1;141(6):1329–1347. doi: 10.1084/jem.141.6.1329

The opsonic fragment of the third component of human complement (C3)

PMCID: PMC2189853  PMID: 236357

Abstract

Human peripheral blood phagocytes ingest Escherichia coli 026:B6 lipopolysaccharide (LPS)-coated paraffin oil droplets containing Oil red O only if fresh serum deposits C3 on the surfaces of the particles (opsonizes them), by reactions involving the properdin system. The rate of binding of purified [125-I]C3 in serum to LPS-coated particles correlated precisely with the rate of acquisition of ingestibility assayed spectrophotometrically. Once opsonized, LPS-coated particles remained fully ingestible and retained fixed [125-I]C3 radioactivity even after exposure to extremes of temperature, pH, ionic strength, phospholipases, urea or guanidine, some nonionic and ionic detergents, and organic solvents. Trypsin, human conglutinogen-activating factor, another heat-stable activity found in human serum, and sodium dodecyl sulfate removed radioactivity and diminished ingestibility of the opsonized particles. Alkylation, reduction plus alkylation and F(ab')2 from anti-C3 blocked ingestibility but did not alter particle-bound radioactivitymelectrophoretic and tryptic peptide autoradiographic analysis of dodecyl sulfate eluates of opsonized particles, cleansed of many contaminating proteins by boiling with 2 M NaCl (yet still opsonized), revealed that the polypeptide with C3-derived radioactivity had a mol wt of approximately 140,000 and was composed of 70,000 mol wt subunits linked by disulfide bonds. Immunochemical analysis and comparison of the peptide structure of the eluate with that of C3 indicated that the opsonic fragment is not the fragment defined as C3b but a smaller derivative of C3.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Abramson N., Alper C. A., Lachmann P. J., Rosen F. S., Jandl J. H. Deficiency of C3 inactivator in man. J Immunol. 1971 Jul;107(1):19–27. [PubMed] [Google Scholar]
  2. Alper C. A., Rosen F. S., Lachmann P. J. Inactivator of the third component of complement as an inhibitor in the properdin pathway. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2910–2913. doi: 10.1073/pnas.69.10.2910. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bokisch V. A., Müller-Eberhard H. J., Cochrane C. G. Isolation of a fragment (C3a) of the third component of human complement containing anaphylatoxin and chemotactic activity and description of an anaphylatoxin inactivator of human serum. J Exp Med. 1969 May 1;129(5):1109–1130. doi: 10.1084/jem.129.5.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Budzko D. B., Bokisch V. A., Müller-Eberhard H. J. A fragment of the third component of human complement with anaphylatoxin activity. Biochemistry. 1971 Mar 30;10(7):1166–1172. doi: 10.1021/bi00783a011. [DOI] [PubMed] [Google Scholar]
  5. Dalmasso A. P., Müller-Eberhard H. J. Physico-chemical characteristics of the third and fourth component of complement after dissociation from complement-cell complexes. Immunology. 1967 Sep;13(3):293–305. [PMC free article] [PubMed] [Google Scholar]
  6. Diamond R. D., May J. E., Kane M. A., Frank M. M., Bennett J. E. The role of the classical and alternate complement pathways in host defenses against Cryptococcus neoformans infection. J Immunol. 1974 Jun;112(6):2260–2270. [PubMed] [Google Scholar]
  7. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  8. Gitlin J. D., Rosen F. S., Lachmann P. J. The mechanism of action of the C3b inactivator (conglutinogen-activating factor) on its naturally occurring substrate, the major fragment of the third component of complement (C3b). J Exp Med. 1975 May 1;141(5):1221–1226. doi: 10.1084/jem.141.5.1221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HIRSCH J. G., STRAUSS B. STUDIES ON HEAT-LABILE OPSONIN IN RABBIT SERUM. J Immunol. 1964 Jan;92:145–154. [PubMed] [Google Scholar]
  10. Jasin H. E. Human heat labile opsonins: evidence for their mediation via the alternate pathway of complement activation. J Immunol. 1972 Jul;109(1):26–31. [PubMed] [Google Scholar]
  11. Johnston R. B., Jr, Klemperer M. R., Alper C. A., Rosen F. S. The enhancement of bacterial phagocytosis by serum. The role of complement components and two cofactors. J Exp Med. 1969 Jun 1;129(6):1275–1290. doi: 10.1084/jem.129.6.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Knüfermann H., Fischer H., Wallach D. F.H. Peptide cleavage in sheep erythrocyte membranes during the action of complement. FEBS Lett. 1971 Aug 15;16(3):167–171. doi: 10.1016/0014-5793(71)80123-0. [DOI] [PubMed] [Google Scholar]
  13. Lachmann P. J., Liske R. The preparation and properties of alexinated intermediates that react with conglutinin. I. Guinea-pig complement. Immunology. 1966 Sep;11(3):243–254. [PMC free article] [PubMed] [Google Scholar]
  14. Lachmann P. J., Müller-Eberhard H. J. The demonstration in human serum of "conglutinogen-activating factor" and its effect on the third component of complement. J Immunol. 1968 Apr;100(4):691–698. [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. NISONOFF A. ENZYMATIC DIGESTION OF RABBIT GAMMA GLOBULIN AND ANTIBODY AND CHROMATOGRAPHY OF DIGESTION PRODUCTS. Methods Med Res. 1964;10:134–141. [PubMed] [Google Scholar]
  19. Nilsson U. R., Mandle R. J., Jr, McConnell-Mapes J. A. Human C3 and C5: subunit structure and modifications by trypsin and C42-C423. J Immunol. 1975 Feb;114(2 Pt 2):815–822. [PubMed] [Google Scholar]
  20. Richards E. G., Chung C. S., Menzel D. B., Olcott H. S. Chromatography of myosin on diethylaminoethyl-Sephadex A-50. Biochemistry. 1967 Feb;6(2):528–540. doi: 10.1021/bi00854a022. [DOI] [PubMed] [Google Scholar]
  21. Ritchie R. F., Alper C. A., Graves J., Pearson N., Larson C. Automated quantitation of proteins in serum and other biologic fluids. Am J Clin Pathol. 1973 Feb;59(2):151–159. doi: 10.1093/ajcp/59.2.151. [DOI] [PubMed] [Google Scholar]
  22. Root R. K., Ellman L., Frank M. M. Bactericidal and opsonic properties of C4-deficient guinea pig serum. J Immunol. 1972 Sep;109(3):477–486. [PubMed] [Google Scholar]
  23. Ruddy S., Austen K. F. C3 inactivator of man. I. Hemolytic measurement by the inactivation of cell-bound C3. J Immunol. 1969 Mar;102(3):533–543. [PubMed] [Google Scholar]
  24. 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]
  25. Shin H. S., Smith M. R., Wood W. B., Jr Heat labile opsonins to pneumococcus. II. Involvement of C3 and C5. J Exp Med. 1969 Dec 1;130(6):1229–1241. doi: 10.1084/jem.130.6.1229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Spitzer R. E., Stitzel A. E., Pauling V. L., Davis N. C., West C. D. The antigenic and molecular alterations of C3 in the fluid phase during an immune reaction in normal human serum. Demonstration of a new conversion product, C3x. J Exp Med. 1971 Sep 1;134(3 Pt 1):656–680. doi: 10.1084/jem.134.3.656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
  28. Stossel T. P., Alper C. A., Rosen F. S. Serum-dependent phagocytosis of paraffin oil emulsified with bacterial lipopolysaccharide. J Exp Med. 1973 Mar 1;137(3):690–705. doi: 10.1084/jem.137.3.690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stossel T. P. Evaluation of opsonic and leukocyte function with a spectrophotometric test in patients with infection and with phagocytic disorders. Blood. 1973 Jul;42(1):121–130. [PubMed] [Google Scholar]
  30. Stossel T. P. Quantitative studies of phagocytosis. Kinetic effects of cations and heat-labile opsonin. J Cell Biol. 1973 Aug;58(2):346–356. doi: 10.1083/jcb.58.2.346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tamura N., Nelson R. A., Jr Three naturally-occurring inhibitors of components of complement in guinea pig and rabbit serum. J Immunol. 1967 Sep;99(3):582–589. [PubMed] [Google Scholar]
  32. Winkelstein J. A., Shin H. S., Wood W. B., Jr Heat labile opsonins to Pneumococcus. 3. The participation of immunoglobulin and of the alternate pathway of C3 activation. J Immunol. 1972 Jun;108(6):1681–1689. [PubMed] [Google Scholar]

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

RESOURCES