Abstract
The membrane attack complex (MAC) of complement was extracted from the membranes of cells lysed by human complement and its properties were compared with those of the fluid phase complex SC5b-9. Upon sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunochemical analysis, the two isolated complexes had identical subunit compositions, except that the MAC lacked the S-protein. The sedimentation coefficient and molecular weight of the extracted and isolated MAC were, respectively, 33.5 S and 1.7 x 10(6) daltons, compared to 23 S and 1.0 x 10(6) dalton for SC5b-9. Because the molecular weight of the MAC is approximately two times greater than that of C5b-0 (800,000 daltons), the MAC is considered the dimer of C5b- 9. Under specified conditions, the 33.5 S dimer could be converted to the 23 S monomer without dissociation of subunits. The MAC had the electron microscopic appearance and dimensions that are characteristic for the complement produced ultrastructural membrane lesions. SC5b-9 had a different ultrastructure that is dissimilar to the morphology of the lesions. The isolated MAC could be reincorporated into phospholipid bilayers and assumed on the surface of the resultant lipid vesicles the orientation and appearance of typical complement lesions.
Full Text
The Full Text of this article is available as a PDF (1.8 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
- BORSOS T., DOURMASHKIN R. R., HUMPHREY J. H. LESIONS IN ERYTHROCYTE MEMBRANES CAUSED BY IMMUNE HAEMOLYSIS. Nature. 1964 Apr 18;202:251–252. doi: 10.1038/202251a0. [DOI] [PubMed] [Google Scholar]
- Brunner J., Skrabal P., Hauser H. Single bilayer vesicles prepared without sonication. Physico-chemical properties. Biochim Biophys Acta. 1976 Dec 2;455(2):322–331. doi: 10.1016/0005-2736(76)90308-4. [DOI] [PubMed] [Google Scholar]
- Dourmashkin R. R. The structural events associated with the attachment of complement components to cell membranes in reactive lysis. Immunology. 1978 Aug;35(2):205–212. [PMC free article] [PubMed] [Google Scholar]
- 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]
- Hesketh T. R., Dourmashkin R. R., Payne S. N., Humphrey J. H., Lachmann P. J. Lesions due to complement in lipid membranes. Nature. 1971 Oct 29;233(5322):620–623. doi: 10.1038/233620a0. [DOI] [PubMed] [Google Scholar]
- Humphrey J. H., Dourmashkin R. R. The lesions in cell membranes caused by complement. Adv Immunol. 1969;11:75–115. doi: 10.1016/s0065-2776(08)60478-2. [DOI] [PubMed] [Google Scholar]
- Klob W. P., Müller-Eberhard H. J. The membrane attack mechanism of complement: the three polypeptide chain structure of the eigth component (C8). J Exp Med. 1976 May 1;143(5):1131–1139. doi: 10.1084/jem.143.5.1131. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kolb W. P., Haxby J. A., Arroyave C. M., Müller-Eberhard H. J. Molecular analysis of the membrane attack mechanism of complement. J Exp Med. 1972 Mar 1;135(3):549–566. doi: 10.1084/jem.135.3.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kolb W. P., Muller-Eberhard H. J. The membrane attack mechanism of complement. Isolation and subunit composition of the C5b-9 complex. J Exp Med. 1975 Apr 1;141(4):724–735. [PMC free article] [PubMed] [Google Scholar]
- Kolb W. P., Müller-Eberhard H. J. The membrane attack mechanism of complement. Verification of a stable C5-9 complex in free solution. J Exp Med. 1973 Aug 1;138(2):438–451. doi: 10.1084/jem.138.2.438. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lachmann P. J., Bowyer D. E., Nicol P., Dawson R. M., Munn E. A. Studies on the terminal stages of complement lysis. Immunology. 1973 Jan;24(1):135–145. [PMC free article] [PubMed] [Google Scholar]
- 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]
- Packman C. H., Rosenfeld S. I., Weed R. I., Leddy J. P. Complement-induced ultrastructural membrane lesions: requirement for terminal components. J Immunol. 1976 Nov;117(5 PT2):1883–1889. [PubMed] [Google Scholar]
- Podack E. R., Kolb W. P., Muller-Eberhard H. J. The C5b-9 complex: subunit composition of the classical and alternative pathway-generated complex. J Immunol. 1976 May;116(5):1431–1434. [PubMed] [Google Scholar]
- Podack E. R., Kolb W. P., Müller-Eberhard H. J. Purification of the sixth and seventh component of human complement without loss of hemolytic activity. J Immunol. 1976 Feb;116(2):263–269. [PubMed] [Google Scholar]
- Podack E. R., Kolb W. P., Müller-Eberhard H. J. The C5b-6 complex: formation, isolation, and inhibition of its activity by lipoprotein and the S-protein of human serum. J Immunol. 1978 Jun;120(6):1841–1848. [PubMed] [Google Scholar]
- Podack E. R., Müller-Eberhard H. J. Binding of desoxycholate, phosphatidylcholine vesicles, lipoprotein and of the S-protein to complexes of terminal complement components. J Immunol. 1978 Sep;121(3):1025–1030. [PubMed] [Google Scholar]
- Tamura N., Shimada A., Chang S. Further evidence for immune cytolysis by antibody and the first eight components of complement in the absence of C9. Immunology. 1972 Jan;22(1):131–140. [PMC free article] [PubMed] [Google Scholar]
- Tranum-Jensen J., Bhakdi S., Bhakdi-Lehnen B., Bjerrum O. J., Speth V. Complement lysis: the ultrastructure and orientation of the C5b-9 complex on target sheep erythrocyte membranes. Scand J Immunol. 1978;7(1):45–46. doi: 10.1111/j.1365-3083.1978.tb00425.x. [DOI] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]