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. 1982 Jan 1;201(1):49–59. doi: 10.1042/bj2010049

The catalytic chain of human complement subcomponent C1r. Purification and N-terminal amino acid sequences of the major cyanogen bromide-cleavage fragments.

G J Arlaud, J Gagnon, R R Porter
PMCID: PMC1163608  PMID: 6282261

Abstract

1. The a- and b-chains of reduced and alkylated human complement subcomponent C1r were separated by high-pressure gel-permeation chromatography and isolated in good yield and in pure form. 2. CNBr cleavage of C1r b-chain yielded eight major peptides, which were purified by gel filtration and high-pressure reversed-phase chromatography. As determined from the sum of their amino acid compositions, these peptides accounted for a minimum molecular weight of 28 000, close to the value 29 100 calculated from the whole b-chain. 3. N-Terminal sequence determinations of C1r b-chain and its CNBr-cleavage peptides allowed the identification of about two-thirds of the amino acids of C1r b-chain. From our results, and on the basis of homology with other serine proteinases, an alignment of the eight CNBr-cleavage peptides from C1r b-chain is proposed. 4. The residues forming the 'charge-relay' system of the active site of serine proteinases (His-57, Asp-102 and Ser-195 in the chymotrypsinogen numbering) are found in the corresponding regions of C1r b-chain, and the amino acid sequence around these residues has been determined. 5. The N-terminal sequence of C1r b-chain has been extended to residue 60 and reveals that C1r b-chain lacks the 'histidine loop', a disulphide bond that is present in all other known serine proteinases.

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Selected References

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

  1. Andrews J. M., Baillie R. D. The enzymatic nature of human c1r: a subcomponent of the first component of complement. J Immunol. 1979 Sep;123(3):1403–1408. [PubMed] [Google Scholar]
  2. Arlaud G. J., Chesne S., Villiers C. L., Colomb M. G. A study on the structure and interactions of the C1 sub-components C1r and C1s in the fluid phase. Biochim Biophys Acta. 1980 Nov 6;616(1):105–115. doi: 10.1016/0005-2744(80)90268-5. [DOI] [PubMed] [Google Scholar]
  3. Arlaud G. J., Sim R. B., Duplaa A. M., Colomb M. G. Differential elution of Clq, Clr and Cls from human Cl bound to immune aggregates. Use in the rapid purification of Cl subcomponents. Mol Immunol. 1979 Jul;16(7):445–450. doi: 10.1016/0161-5890(79)90069-5. [DOI] [PubMed] [Google Scholar]
  4. Arlaud G. J., Villiers C. L., Chesne S., Colomb M. G. Purified proenzyme C1r. Some characteristics of its activation and subsequent proteolytic cleavage. Biochim Biophys Acta. 1980 Nov 6;616(1):116–129. doi: 10.1016/0005-2744(80)90269-7. [DOI] [PubMed] [Google Scholar]
  5. Assimeh S. N., Chapuis R. M., Isliker H. Studies on the precursor form of the first component of complement. II. Proteolytic fragmentation of Clr. Immunochemistry. 1978 Jan;15(1):13–17. doi: 10.1016/0161-5890(78)90020-2. [DOI] [PubMed] [Google Scholar]
  6. Bridgen P. J., Cross G. A., Bridgen J. N-terminal amino acid sequences of variant-specific surface antigens from Trypanosoma brucei. Nature. 1976 Oct 14;263(5578):613–614. doi: 10.1038/263613a0. [DOI] [PubMed] [Google Scholar]
  7. De Bracco M. M., Stroud R. M. C1r, subunit of the first complement component: purification, properties, and assay based on its linking role. J Clin Invest. 1971 Apr;50(4):838–848. doi: 10.1172/JCI106555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dodds A. W., Sim R. B., Porter R. R., Kerr M. A. Activation of the first component of human complement (C1) by antibody-antigen aggregates. Biochem J. 1978 Nov 1;175(2):383–390. doi: 10.1042/bj1750383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Jackson C. M., Nemerson Y. Blood coagulation. Annu Rev Biochem. 1980;49:765–811. doi: 10.1146/annurev.bi.49.070180.004001. [DOI] [PubMed] [Google Scholar]
  11. Johnson D. M., Gagnon J., Reid K. B. Factor D of the alternative pathway of human complement. Purification, alignment and N-terminal amino acid sequences of the major cyanogen bromide fragments, and localization of the serine residue at the active site. Biochem J. 1980 Jun 1;187(3):863–874. doi: 10.1042/bj1870863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kerlavage A. R., Taylor S. S. Covalent modification of an adenosine 3':5'-monophosphate binding site of the regulatory subunit of cAMP-dependent protein kinase II with 8-azidoadenosine 3':5'-monophosphate. Identification of a single modified tyrosine residue. J Biol Chem. 1980 Sep 25;255(18):8483–8488. [PubMed] [Google Scholar]
  13. LEPOW I. H., NAFF G. B., TODD E. W., PENSKY J., HINZ C. F. Chromatographic resolution of the first component of human complement into three activities. J Exp Med. 1963 Jun 1;117:983–1008. doi: 10.1084/jem.117.6.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Naff G. B., Ratnoff O. S. The enzymatic nature of C'1r. Conversion of C'1s to C'1 esterase and digestion of amino acid esters by C'1r. J Exp Med. 1968 Oct 1;128(4):571–593. doi: 10.1084/jem.128.4.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Porter R. R. The eleventh Hopkins Memorial Lecture: The biochemistry of complement. Biochem Soc Trans. 1977;5(6):1659–1674. doi: 10.1042/bst0051659. [DOI] [PubMed] [Google Scholar]
  17. Sim R. B., Porter R. R. Isolation and comparison of the proenzyme and activated forms of the human serum complement subcomponents C1r and C1s. Biochem Soc Trans. 1976;4(1):127–129. doi: 10.1042/bst0040127. [DOI] [PubMed] [Google Scholar]
  18. Sim R. B., Porter R. R., Reid K. B., Gigli I. The structure and enzymic activities of the C1r and C1s subcomponents of C1, the first component of human serum complement. Biochem J. 1977 May 1;163(2):219–227. doi: 10.1042/bj1630219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Takahashi K., Nagasawa S., Koyama J. A gross structure of an activated form of a subunit of the first component of human complement. Clr. FEBS Lett. 1975 Jul 15;55(1):156–160. doi: 10.1016/0014-5793(75)80982-3. [DOI] [PubMed] [Google Scholar]
  20. Valet G., Cooper N. R. Isolation and characterization of the proenzyme form of the C1r subunit of the first complement component. J Immunol. 1974 May;112(5):1667–1673. [PubMed] [Google Scholar]
  21. Volanakis J. E., Schrohenloher R. E., Stroud R. M. Human factor D of the alternative complement pathway: purification and characterization. J Immunol. 1977 Jul;119(1):337–342. [PubMed] [Google Scholar]
  22. Woods K. R., Wang K. T. Separation of dansyl-amino acids by polyamide layer chromatography. Biochim Biophys Acta. 1967 Feb 21;133(2):369–370. doi: 10.1016/0005-2795(67)90078-5. [DOI] [PubMed] [Google Scholar]
  23. Ziccardi R. J., Cooper N. R. Activation of C1r by proteolytic cleavage. J Immunol. 1976 Feb;116(2):504–509. [PubMed] [Google Scholar]
  24. Ziccardi R. J., Cooper N. R. Physicochemical and functional characterization of the C1r subunit of the first complement component. J Immunol. 1976 Feb;116(2):496–503. [PubMed] [Google Scholar]
  25. de Haën C., Neurath H., Teller D. C. The phylogeny of trypsin-related serine proteases and their zymogens. New methods for the investigation of distant evolutionary relationships. J Mol Biol. 1975 Feb 25;92(2):225–259. doi: 10.1016/0022-2836(75)90225-9. [DOI] [PubMed] [Google Scholar]

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