Abstract
The ultrastructure of the membrane attack complex (MAC) of complement had been described as representing a hollow cylinder of defined dimensions that is composed of the proteins C5b, C6, C7, C8, and C9. After the characteristic cylindrical structure was identified as polymerized C9 [poly(C9)], the question arose as to the ultrastructural identity and topology of the C9-polymerizing complex C5b-8. An electron microscopic analysis of isolated MAC revealed an asymmetry of individual complexes with respect to their length. Whereas the length of one boundary (+/- SEM) was always 16 +/- 1 nm, the length of the other varied between 16 and 32 nm. In contrast, poly(C9), formed spontaneously from isolated C9, had a uniform tubule length (+/- SEM) of 16 +/- 1 nm. On examination of MAC-phospholipid vesicle complexes, an elongated structure was detected that was closely associated with the poly(C9) tubule and that extended 16-18 nm beyond the torus of the tubule and 28-30 nm above the membrane surface. The width of this structure varied depending on its two-dimensional projection in the electron microscope. By using biotinyl C5b-6 in the formation of the MAC and avidin-coated colloidal gold particles for the ultrastructural analysis, this heretofore unrecognized subunit of the MAC could be identified as the tetramolecular C5b-8 complex. Identification also was achieved by using anti-C5 Fab-coated colloidal gold particles. A similar elongated structure of 25 nm length (above the surface of the membrane) was observed on single C5b-8-vesicle complexes. It is concluded that the C5b-8 complex, which catalyzes poly(C9) formation, constitutes a structure of discrete morphology that remains as such identifiable in the fully assembled MAC, in which it is closely associated with the poly(C9) tubule.
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- 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]
- Bhakdi S., Ey P., Bhakdi-Lehnen B. Isolation of the terminal complement complex from target sheep erythrocyte membranes. Biochim Biophys Acta. 1976 Feb 6;419(3):445–457. doi: 10.1016/0005-2736(76)90258-3. [DOI] [PubMed] [Google Scholar]
- Bhakdi S., Tranum-Jensen J. Molecular nature of the complement lesion. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5655–5659. doi: 10.1073/pnas.75.11.5655. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Biesecker G., Müller-Eberhard H. J. The ninth component of human complement: purification and physicochemical characterization. J Immunol. 1980 Mar;124(3):1291–1296. [PubMed] [Google Scholar]
- Biesecker G., Podack E. R., Halverson C. A., Müller-Eberhard H. J. C5b-9 dimer: isolation from complement lysed cells and ultrastructural identification with complement-dependent membrane lesions. J Exp Med. 1979 Feb 1;149(2):448–458. doi: 10.1084/jem.149.2.448. [DOI] [PMC free article] [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]
- Esser A. F., Kolb W. P., Podack E. R., Müller-Eberhard H. J. Molecular reorganization of lipid bilayers by complement: a possible mechanism for membranolysis. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1410–1414. doi: 10.1073/pnas.76.3.1410. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horisberger M., Rosset J., Bauer H. Colloidal gold granules as markers for cell surface receptors in the scanning electron microscope. Experientia. 1975 Oct 15;31(10):1147–1149. doi: 10.1007/BF02326761. [DOI] [PubMed] [Google Scholar]
- Hu V. W., Esser A. F., Podack E. R., Wisnieski B. J. The membrane attack mechanism of complement: photolabeling reveals insertion of terminal proteins into target membrane. J Immunol. 1981 Jul;127(1):380–386. [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]
- Mayer M. M. Complement, past and present. Harvey Lect. 1978;72:139–193. [PubMed] [Google Scholar]
- Mayer M. M. Mechanism of cytolysis by complement. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2954–2958. doi: 10.1073/pnas.69.10.2954. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Podack E. R., Esser A. F., Biesecker G., Müller-Eberhard H. J. Membrane attack complex of complement: a structural analysis of its assembly. J Exp Med. 1980 Feb 1;151(2):301–313. doi: 10.1084/jem.151.2.301. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Podack E. R., Kolb W. P., Esser A. F., Müller-Eberhard H. J. Structural similarities between C6 and C7 of human complement. J Immunol. 1979 Sep;123(3):1071–1077. [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]
- Podack E. R., Müller-Eberhard H. J., Horst H., Hoppe W. Membrane attach complex of complement (MAC): three-dimensional analysis of MAC-phospholipid vesicle recombinants. J Immunol. 1982 May;128(5):2353–2357. [PubMed] [Google Scholar]
- Podack E. R., Müller-Eberhard H. J. Limited proteolysis of C5b-6: functional stability of the degraded complex. J Immunol. 1980 Jan;124(1):332–336. [PubMed] [Google Scholar]
- Podack E. R., Müller-Eberhard H. J. Membrane attack complex of complement. Evidence for its dimeric structure based on hybrid formation. J Biol Chem. 1981 Apr 10;256(7):3145–3148. [PubMed] [Google Scholar]
- Podack E. R., Stoffel W., Esser A. F., Müller-Eberhard H. J. Membrane attack complex of complement: distribution of subunits between the hydrocarbon phase of target membranes and water. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4544–4548. doi: 10.1073/pnas.78.7.4544. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Podack E. R., Tschoop J., Müller-Eberhard H. J. Molecular organization of C9 within the membrane attack complex of complement. Induction of circular C9 polymerization by the C5b-8 assembly. J Exp Med. 1982 Jul 1;156(1):268–282. doi: 10.1084/jem.156.1.268. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Podack E. R., Tschopp J. Polymerization of the ninth component of complement (C9): formation of poly(C9) with a tubular ultrastructure resembling the membrane attack complex of complement. Proc Natl Acad Sci U S A. 1982 Jan;79(2):574–578. doi: 10.1073/pnas.79.2.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sundsmo J. S., Müller-Eberhard H. J. Neoantigen of the complement membrane attack complex of cytotoxic human peripheral blood lymphocytes. J Immunol. 1979 Jun;122(6):2371–2378. [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]
- Tschopp J., Müller-Eberhard H. J., Podack E. R. Formation of transmembrane tubules by spontaneous polymerization of the hydrophilic complement protein C9. Nature. 1982 Aug 5;298(5874):534–538. doi: 10.1038/298534a0. [DOI] [PubMed] [Google Scholar]
- Tschopp J., Podack E. R. Membranolysis by the ninth component of human complement. Biochem Biophys Res Commun. 1981 Jun 16;100(3):1409–1414. doi: 10.1016/0006-291x(81)91981-1. [DOI] [PubMed] [Google Scholar]