Skip to main content
PMC home page
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 1995 Nov;4(11):2433–2435. doi: 10.1002/pro.5560041123

Identification of the catalytic histidine residue participating in the charge-relay system of carboxypeptidase Y.

G Jung 1, H Ueno 1, R Hayashi 1, T H Liao 1
PMCID: PMC2143012  PMID: 8563642

Abstract

The essential histidine residue of carboxypeptidase Y (CPY) was modified by a site-specific reagent, a chloromethylketone derivative of benzyloxycarbonyl-L-phenylalanine. The single modified histidine residue was converted to N tau-carboxy-methyl histidine (cmHis) upon performic acid oxidation. A peptide containing cmHis was isolated from the tryptic-thermolytic digest. Based on the amino acid composition and sequence analysis, the peptide is shown to be Val-Phe-Asp-Gly-Gly-cmHis-MetO2-Val-Pro, which was derived from CPY cleaved by trypsin at Arg 391 and thermolysin at Phe 401, and thus His 397 was modified. This histidine residue has been implicated previously by X-ray analysis to participate in the charge-relay system of CPY.

Full Text

The Full Text of this article is available as a PDF (259.0 KB).

Selected References

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

  1. Bech L. M., Breddam K. Inactivation of carboxypeptidase Y by mutational removal of the putative essential histidyl residue. Carlsberg Res Commun. 1989;54(5):165–171. doi: 10.1007/BF02904470. [DOI] [PubMed] [Google Scholar]
  2. Blow D. M., Birktoft J. J., Hartley B. S. Role of a buried acid group in the mechanism of action of chymotrypsin. Nature. 1969 Jan 25;221(5178):337–340. doi: 10.1038/221337a0. [DOI] [PubMed] [Google Scholar]
  3. DeTar D. F., Rogers F. F., Jr, Bach H. Sequence peptide polymers. 3. Poly Asp(OH)-Ser(H)-Gly and other serine-containing polymers. J Am Chem Soc. 1967 Jun 7;89(12):3039–3045. doi: 10.1021/ja00988a043. [DOI] [PubMed] [Google Scholar]
  4. Endrizzi J. A., Breddam K., Remington S. J. 2.8-A structure of yeast serine carboxypeptidase. Biochemistry. 1994 Sep 20;33(37):11106–11120. doi: 10.1021/bi00203a007. [DOI] [PubMed] [Google Scholar]
  5. Greene L. J., Bartelt D. C. Specific hydrolysis by trypsin at alkaline pH. Methods Enzymol. 1977;47:170–174. doi: 10.1016/0076-6879(77)47021-6. [DOI] [PubMed] [Google Scholar]
  6. HIRS C. H. The oxidation of ribonuclease with performic acid. J Biol Chem. 1956 Apr;219(2):611–621. [PubMed] [Google Scholar]
  7. Hayashi R., Bai Y., Hata T. Further confirmation of carboxypeptidase Y as a metal-free enzyme having a reactive serine residue. J Biochem. 1975 Jun;77(6):1313–1318. [PubMed] [Google Scholar]
  8. Hayashi R. Carboxypeptidase Y. Methods Enzymol. 1976;45:568–587. doi: 10.1016/s0076-6879(76)45051-6. [DOI] [PubMed] [Google Scholar]
  9. Hayashi R., Moore S., Stein W. H. Carboxypeptidase from yeast. Large scale preparation and the application to COOH-terminal analysis of peptides and proteins. J Biol Chem. 1973 Apr 10;248(7):2296–2302. [PubMed] [Google Scholar]
  10. Hayashi R., Moore S., Stein W. H. Serine at the active center of yeast carboxypeptidase. J Biol Chem. 1973 Dec 25;248(24):8366–8369. [PubMed] [Google Scholar]
  11. Kuhn R. W., Walsh K. A., Neurath H. Reaction of yeast carboxypeptidase C1 with group-specific reagents. Biochemistry. 1976 Nov 2;15(22):4881–4885. doi: 10.1021/bi00667a020. [DOI] [PubMed] [Google Scholar]
  12. Liao T. H., Salnikow J., Moore S., Stein W. H. Bovine pancreatic deoxyribonuclease A. Isolation of cyanogen bromide peptides; complete covalent structure of the polypeptide chain. J Biol Chem. 1973 Feb 25;248(4):1489–1495. [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES