Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1989 May 15;260(1):259–263. doi: 10.1042/bj2600259

A continuous fluorimetric assay for clostridial collagenase and Pz-peptidase activity.

A J Barrett 1, C G Knight 1, M A Brown 1, U Tisljar 1
PMCID: PMC1138654  PMID: 2549953

Abstract

The peptide derivative N alpha-(2,4-dinitrophenyl)-L-prolyl-L-leucyl-glycyl-L-prolyl-L-tryptophanyl-D- lysine (Dnp-Pro-Leu-Gly-Pro-Trp-D-Lys) has been found to be a convenient substrate for the assay of clostridial collagenase and Pz-peptidase. The substrate shows a 25-fold enhancement of fluorescence (gamma ex. 283 nm, lambda em. 350 nm) following hydrolysis of the Leu2-Gly3 peptide bond. The value of Km for clostridial collagenase was 17 microM. The substrate for the first time makes possible continuous fluorimetric assays for Pz-peptidase and clostridial collagenase.

Full text

PDF
259

Selected References

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

  1. Chikuma T., Ishii Y., Kato T. Highly sensitive assay for PZ-peptidase activity by high-performance liquid chromatography. J Chromatogr. 1985 Nov 27;348(1):205–212. doi: 10.1016/s0021-9673(01)92454-9. [DOI] [PubMed] [Google Scholar]
  2. Hino M., Nagatsu T. Separation of two PZ-peptidases from bovine dental follicle. Biochim Biophys Acta. 1976 Apr 8;429(2):555–563. doi: 10.1016/0005-2744(76)90303-x. [DOI] [PubMed] [Google Scholar]
  3. Jackson G. E., Young N. M. The dinitrophenyl group as a selective label in high-performance liquid chromatography of peptides. Anal Biochem. 1987 Apr;162(1):251–256. doi: 10.1016/0003-2697(87)90034-0. [DOI] [PubMed] [Google Scholar]
  4. Lakowicz J. R., Balter A. Direct recording of the initially excited and the solvent relaxed fluorescence emission spectra of tryptophan by phase sensitive detection of fluorescence. Photochem Photobiol. 1982 Aug;36(2):125–132. doi: 10.1111/j.1751-1097.1982.tb04353.x. [DOI] [PubMed] [Google Scholar]
  5. Lessley B. A., Garner D. L. Identification and distribution of Pz-peptidases A and B in human semen. J Androl. 1985 Nov-Dec;6(6):372–378. doi: 10.1002/j.1939-4640.1985.tb03296.x. [DOI] [PubMed] [Google Scholar]
  6. Morales T. I., Woessner J. F., Jr PZ-peptidase from chick embryos. Purification, properties, and action on collagen peptides. J Biol Chem. 1977 Jul 25;252(14):4855–4860. [PubMed] [Google Scholar]
  7. Porter R. R., Sanger F. The free amino groups of haemoglobins. Biochem J. 1948;42(2):287–294. doi: 10.1042/bj0420287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Rajabi M., Woessner J. F., Jr Rise in serum levels of PZ-peptidase, an enzyme involved in collagen breakdown, in human pregnancy and labor. Am J Obstet Gynecol. 1984 Dec 1;150(7):821–826. doi: 10.1016/0002-9378(84)90456-3. [DOI] [PubMed] [Google Scholar]
  9. Steinbrink D. R., Bond M. D., Van Wart H. E. Substrate specificity of beta-collagenase from Clostridium histolyticum. J Biol Chem. 1985 Mar 10;260(5):2771–2776. [PubMed] [Google Scholar]
  10. Stryer L. Fluorescence energy transfer as a spectroscopic ruler. Annu Rev Biochem. 1978;47:819–846. doi: 10.1146/annurev.bi.47.070178.004131. [DOI] [PubMed] [Google Scholar]
  11. WUENSCH E., HEIDRICH H. G. ZUR QUANTITATIVEN BESTIMMUNG DER KOLLAGENASE. Hoppe Seylers Z Physiol Chem. 1963;333:149–151. doi: 10.1515/bchm2.1963.333.1.149. [DOI] [PubMed] [Google Scholar]
  12. Yaron A., Carmel A., Katchalski-Katzir E. Intramolecularly quenched fluorogenic substrates for hydrolytic enzymes. Anal Biochem. 1979 May;95(1):228–235. doi: 10.1016/0003-2697(79)90210-0. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES