Skip to main content
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1992 May;88(2):368–372. doi: 10.1111/j.1365-2249.1992.tb03090.x

Functional activities of synthetic anaphylatoxic peptides in widely used biological assays.

A Kola 1, A Klos 1, W Bautsch 1, T Kretzschmar 1, J Köhl 1
PMCID: PMC1554312  PMID: 1572104

Abstract

A comparison study was carried out between the modern ATP release assay (ARA) with guinea-pig platelets and the traditional guinea-pig ileum contraction assay (ICA). The biological activities of the anaphylatoxin C3a and synthetic C3a analogue peptides were determined in both assays. In dose-response curves with C3a, a human C3a peptide with the last 21 amino acids of the C terminus (C3a 56-77) and a peptide with 13 amino acids which was acylated N-terminal with the aromatic fluorenylmethoxycarbonyl group and an aminohexanoyl group (Fmoc-Ahx YRRGRAAALGLAR) were tested. The ARA turned out to be 100 times more sensitive than the ICA. In contrast to previous reports the 21 amino acid long C3a analogue peptide did not exhibit full C3a activity but only 7% (ARA) or 12% (ICA). The potentiation of biological activity in the ARA by coupling non-peptide acyl-residues N terminal to peptide C3a analogues could be confirmed with Fmoc-Ahx-YRRGRAAALGLAR in the ICA. In addition, the tri-peptide Fmoc-Ahx-LAR displayed C3a specific activity in the ICA demonstrated by desensitization experiments.

Full text

PDF
368

Selected References

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

  1. ASTER R. H., JANDL J. H. PLATELET SEQUESTRATION IN MAN. I. METHODS. J Clin Invest. 1964 May;43:843–855. doi: 10.1172/JCI104970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Becker S., Meuer S., Hadding U., Bitter-Suermann D. Platelet activation: a new biological activity of guinea-pig C3a anaphylatoxin. Scand J Immunol. 1978;7(2):173–180. doi: 10.1111/j.1365-3083.1978.tb00441.x. [DOI] [PubMed] [Google Scholar]
  3. Bitter-Suermann D., Becker S., Meuer S., Schorlemmer H. U., Hadding U., Andreatta R. Comparative study on biological effects of the guinea pig complement-peptide C3a and C3a-related synthetic oligopeptides. Mol Immunol. 1980 Oct;17(10):1257–1261. doi: 10.1016/0161-5890(80)90022-x. [DOI] [PubMed] [Google Scholar]
  4. Bitter-Suermann D., Burger R. Guinea pigs deficient in C2, C4, C3 or the C3a receptor. Prog Allergy. 1986;39:134–158. [PubMed] [Google Scholar]
  5. Caporale L. H., Tippett P. S., Erickson B. W., Hugli T. E. The active site of C3a anaphylatoxin. J Biol Chem. 1980 Nov 25;255(22):10758–10763. [PubMed] [Google Scholar]
  6. Cochrane C. G., Müller-Eberhard H. J. The derivation of two distinct anaphylatoxin activities from the third and fifth components of human complement. J Exp Med. 1968 Feb 1;127(2):371–386. doi: 10.1084/jem.127.2.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fukuoka Y., Hugli T. E. Anaphylatoxin binding and degradation by rat peritoneal mast cells. Mechanisms of degranulation and control. J Immunol. 1990 Sep 15;145(6):1851–1858. [PubMed] [Google Scholar]
  8. Fukuoka Y., Hugli T. E. Demonstration of a specific C3a receptor on guinea pig platelets. J Immunol. 1988 May 15;140(10):3496–3501. [PubMed] [Google Scholar]
  9. Gerardy-Schahn R., Ambrosius D., Casaretto M., Grötzinger J., Saunders D., Wollmer A., Brandenburg D., Bitter-Suermann D. Design and biological activity of a new generation of synthetic C3a analogues by combination of peptidic and non-peptidic elements. Biochem J. 1988 Oct 1;255(1):209–216. [PMC free article] [PubMed] [Google Scholar]
  10. Gerardy-Schahn R., Ambrosius D., Saunders D., Casaretto M., Mittler C., Karwarth G., Görgen S., Bitter-Suermann D. Characterization of C3a receptor-proteins on guinea pig platelets and human polymorphonuclear leukocytes. Eur J Immunol. 1989 Jun;19(6):1095–1102. doi: 10.1002/eji.1830190620. [DOI] [PubMed] [Google Scholar]
  11. Hugli T. E., Erickson B. W. Synthetic peptides with the biological activities and specificity of human C3a anaphylatoxin. Proc Natl Acad Sci U S A. 1977 May;74(5):1826–1830. doi: 10.1073/pnas.74.5.1826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hugli T. E., Müller-Eberhard H. J. Anaphylatoxins: C3a and C5a. Adv Immunol. 1978;26:1–53. doi: 10.1016/s0065-2776(08)60228-x. [DOI] [PubMed] [Google Scholar]
  13. Hugli T. E. Structure and function of C3a anaphylatoxin. Curr Top Microbiol Immunol. 1990;153:181–208. doi: 10.1007/978-3-642-74977-3_10. [DOI] [PubMed] [Google Scholar]
  14. Kretzschmar T., Kahl K., Rech K., Bautsch W., Köhl J., Bitter-Suermann D. Characterization of the C5a receptor on guinea pig platelets. Immunobiology. 1991 Nov;183(5):418–432. doi: 10.1016/S0171-2985(11)80526-7. [DOI] [PubMed] [Google Scholar]
  15. Köhl J., Casaretto M., Gier M., Karwath G., Gietz C., Bautsch W., Saunders D., Bitter-Suermann D. Reevaluation of the C3a active site using short synthetic C3a analogues. Eur J Immunol. 1990 Jul;20(7):1463–1468. doi: 10.1002/eji.1830200709. [DOI] [PubMed] [Google Scholar]
  16. Lu Z. X., Fok K. F., Erickson B. W., Hugli T. E. Conformational analysis of COOH-terminal segments of human C3a. Evidence of ordered conformation in an active 21-residue peptide. J Biol Chem. 1984 Jun 25;259(12):7367–7370. [PubMed] [Google Scholar]
  17. Zanker B., Rasokat H., Hadding U., Bitter-Suermann D. C3a induced activation and stimulus specific reversible desensitization of guinea pig platelets. Agents Actions Suppl. 1982;11:147–157. [PubMed] [Google Scholar]
  18. del Balzo U., Polley M. J., Levi R. C3a-induced contraction of guinea pig ileum consists of two components: fast histamine-mediated and slow prostanoid-mediated. J Pharmacol Exp Ther. 1989 Mar;248(3):1003–1009. [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES