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. 1986 Nov 1;239(3):763–767. doi: 10.1042/bj2390763

Phenotyping of human complement component C4, a class-III HLA antigen.

E Sim, S J Cross
PMCID: PMC1147352  PMID: 3103606

Abstract

The plasma complement protein C4 is encoded at two highly polymorphic loci, A and B, within the class-III region of the major histocompatibility complex. At least 34 different polymorphic variants of human C4 have been identified, including non-expressed or 'null' alleles. The main method of identification of C4 polymorphic allotypes is separation on the basis of charge by agarose-gel electrophoresis of plasma. On staining by immunofixation with anti-C4 antibodies, each C4 type gives three major bands, but, since individuals can have up to five allotypes, the overlapping banding pattern is difficult to interpret. We show that digestion of plasma samples with carboxypeptidase B, which removes C-terminal basic amino acids, before electrophoresis, produces a single, sharp, distinct band for each allotype and allows identification of the biochemical basis of the multiple banding pattern previously observed in C4 phenotype determination.

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Selected References

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

  1. Awdeh Z. L., Alper C. A., Eynon E., Alosco S. M., Stein R., Yunis E. J. Unrelated individuals matched for MHC extended haplotypes and HLA-identical siblings show comparable responses in mixed lymphocyte culture. Lancet. 1985 Oct 19;2(8460):853–856. doi: 10.1016/s0140-6736(85)90123-0. [DOI] [PubMed] [Google Scholar]
  2. Awdeh Z. L., Alper C. A. Inherited structural polymorphism of the fourth component of human complement. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3576–3580. doi: 10.1073/pnas.77.6.3576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Awdeh Z. L., Raum D., Alper C. A. Genetic polymorphism of human complement C4 and detection of heterozygotes. Nature. 1979 Nov 8;282(5735):205–207. doi: 10.1038/282205a0. [DOI] [PubMed] [Google Scholar]
  4. Belt K. T., Carroll M. C., Porter R. R. The structural basis of the multiple forms of human complement component C4. Cell. 1984 Apr;36(4):907–914. doi: 10.1016/0092-8674(84)90040-0. [DOI] [PubMed] [Google Scholar]
  5. Campbell R. D., Gagnon J., Porter R. R. Amino acid sequence around the thiol and reactive acyl groups of human complement component C4. Biochem J. 1981 Nov 1;199(2):359–370. doi: 10.1042/bj1990359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carroll M. C., Belt T., Palsdottir A., Porter R. R. Structure and organization of the C4 genes. Philos Trans R Soc Lond B Biol Sci. 1984 Sep 6;306(1129):379–388. doi: 10.1098/rstb.1984.0098. [DOI] [PubMed] [Google Scholar]
  7. Carroll M. C., Palsdottir A., Belt K. T., Porter R. R. Deletion of complement C4 and steroid 21-hydroxylase genes in the HLA class III region. EMBO J. 1985 Oct;4(10):2547–2552. doi: 10.1002/j.1460-2075.1985.tb03969.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dawkins R. L., Christiansen F. T., Kay P. H., Garlepp M., McCluskey J., Hollingsworth P. N., Zilko P. J. Disease associations with complotypes, supratypes and haplotypes. Immunol Rev. 1983;70:5–22. doi: 10.1111/j.1600-065x.1983.tb00707.x. [DOI] [PubMed] [Google Scholar]
  9. FOLK J. E., PIEZ K. A., CARROLL W. R., GLADNER J. A. Carboxy-peptidase B. 4. Purification and characterization of the porcine enzyme. J Biol Chem. 1960 Aug;235:2272–2277. [PubMed] [Google Scholar]
  10. Fernandez H. N., Hugli T. E. Primary structural analysis of the polypeptide portion of human C5a anaphylatoxin. Polypeptide sequence determination and assignment of the oligosaccharide attachment site in C5a. J Biol Chem. 1978 Oct 10;253(19):6955–6964. [PubMed] [Google Scholar]
  11. Fielder A. H., Walport M. J., Batchelor J. R., Rynes R. I., Black C. M., Dodi I. A., Hughes G. R. Family study of the major histocompatibility complex in patients with systemic lupus erythematosus: importance of null alleles of C4A and C4B in determining disease susceptibility. Br Med J (Clin Res Ed) 1983 Feb 5;286(6363):425–428. doi: 10.1136/bmj.286.6363.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Isenman D. E., Young J. R. Covalent binding properties of the C4A and C4B isotypes of the fourth component of human complement on several C1-bearing cell surfaces. J Immunol. 1986 Apr 1;136(7):2542–2550. [PubMed] [Google Scholar]
  13. Isenman D. E., Young J. R. The molecular basis for the difference in immune hemolysis activity of the Chido and Rodgers isotypes of human complement component C4. J Immunol. 1984 Jun;132(6):3019–3027. [PubMed] [Google Scholar]
  14. Law S. K., Dodds A. W., Porter R. R. A comparison of the properties of two classes, C4A and C4B, of the human complement component C4. EMBO J. 1984 Aug;3(8):1819–1823. doi: 10.1002/j.1460-2075.1984.tb02052.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Law S. K., Gagnon J. The primary structure of the fourth component of human complement (C4)-C-terminal peptides. Biosci Rep. 1985 Oct-Nov;5(10-11):913–921. doi: 10.1007/BF01119904. [DOI] [PubMed] [Google Scholar]
  16. Mauff G., Alper C. A., Awdeh Z., Batchelor J. R., Bertrams J., Bruun-Petersen G., Dawkins R. L., Démant P., Edwards J., Grosse-Wilde H. Statement on the nomenclature of human C4 allotypes. Immunobiology. 1983 Mar;164(2):184–191. doi: 10.1016/s0171-2985(83)80009-6. [DOI] [PubMed] [Google Scholar]
  17. McKay T. J., Phelan A. W., Plummer T. H., Jr Comparative studies on human carboxypeptidases B and N. Arch Biochem Biophys. 1979 Oct 15;197(2):487–492. doi: 10.1016/0003-9861(79)90271-6. [DOI] [PubMed] [Google Scholar]
  18. O'Neill G. J., Nerl C. W., Kay P. H., Christiansen F. T., McCluskey J., Dawkins R. L. Complement C4 is a marker for adult rheumatoid arthritis. Lancet. 1982 Jul 24;2(8291):214–214. doi: 10.1016/s0140-6736(82)91057-1. [DOI] [PubMed] [Google Scholar]
  19. Palsdottir A., Cross S. J., Edwards J. H., Carroll M. C. Correlation between a DNA restriction fragment length polymorphism and C4A6 protein. Nature. 1983 Dec 8;306(5943):615–616. doi: 10.1038/306615a0. [DOI] [PubMed] [Google Scholar]
  20. Perryman M. B., Knell J. D., Roberts R. Carboxypeptidase-catalyzed hydrolysis of C-terminal lysine: mechanism for in vivo production of multiple forms of creatine kinase in plasma. Clin Chem. 1984 May;30(5):662–664. [PubMed] [Google Scholar]
  21. Porter R. R. Complement polymorphism, the major histocompatibility complex and associated diseases: a speculation. Mol Biol Med. 1983 Jul;1(1):161–168. [PubMed] [Google Scholar]
  22. Porter R. R. The complement components coded in the major histocompatibility complexes and their biological activities. Immunol Rev. 1985 Oct;87:7–17. doi: 10.1111/j.1600-065x.1985.tb01142.x. [DOI] [PubMed] [Google Scholar]
  23. Roos M. H., Mollenhauer E., Démant P., Rittner C. A molecular basis for the two locus model of human complement component C4. Nature. 1982 Aug 26;298(5877):854–856. doi: 10.1038/298854a0. [DOI] [PubMed] [Google Scholar]
  24. Schendel D. J., O'Neill G. J., Wank R. MHC-linked class III genes. Analysis of C4 gene frequencies, complotypes and associations with distinct HLA haplotypes in German Caucasians. Immunogenetics. 1984;20(1):23–31. doi: 10.1007/BF00373444. [DOI] [PubMed] [Google Scholar]
  25. Shreffler D. C., Atkinson J. P., Chan A. C., Karp D. R., Killion C. C., Ogata R. T., Rosa P. A. The C4 and Slp genes of the complement region of the murine H-2 major histocompatibility complex. Philos Trans R Soc Lond B Biol Sci. 1984 Sep 6;306(1129):395–403. doi: 10.1098/rstb.1984.0100. [DOI] [PubMed] [Google Scholar]
  26. Sim E., Law S. K. Hydralazine binds covalently to complement component C4. Different reactivity of C4A and C4B gene products. FEBS Lett. 1985 May 20;184(2):323–327. doi: 10.1016/0014-5793(85)80631-1. [DOI] [PubMed] [Google Scholar]
  27. Sim R. B., Sim E. Autolytic fragmentation of complement components C3 and C4 under denaturing conditions, a property shared with alpha 2-macroglobulin. Biochem J. 1981 Jan 1;193(1):129–141. doi: 10.1042/bj1930129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tilley C. A., Romans D. G., Crookston M. C. Localisation of Chido and Rodgers determinants to the C4d fragment of human C4. Nature. 1978 Dec 14;276(5689):713–715. doi: 10.1038/276713a0. [DOI] [PubMed] [Google Scholar]
  29. Whitehead A. S., Woods D. E., Fleischnick E., Chin J. E., Yunis E. J., Katz A. J., Gerald P. S., Alper C. A., Colten H. R. DNA polymorphism of the C4 genes. A new marker for analysis of the major histocompatibility complex. N Engl J Med. 1984 Jan 12;310(2):88–91. doi: 10.1056/NEJM198401123100204. [DOI] [PubMed] [Google Scholar]

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