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. 1969 Aug;63(4):1389–1394. doi: 10.1073/pnas.63.4.1389

THE AMINO ACID SEQUENCE OF BOVINE CARBOXYPEPTIDASE A*

Ralph A Bradshaw 1,†, Lowell H Ericsson 1, Kenneth A Walsh 1, Hans Neurath 1
PMCID: PMC223477  PMID: 5260942

Abstract

The amino acid sequence of the four fragments produced by treatment of bovine carboxypeptidase A with cyanogen bromide has been completed. The alignment of these fragments, previously established by peptic digest of the whole protein, allows for the description of the complete primary structure of the molecule. A comparison of the proposed functional residues, identified by X-ray diffraction analyses, has either confirmed their assignments or provided their correct identity. The functional and structural residues of principal importance include Arg 145, Tyr 248, and Glu 270 as the binding site of the substrate carboxyl group, the proton donor, and the nucleophilic moiety, respectively, which were correctly assigned; His 196 as the third zinc ligand and Tyr 265 as the binding site of the α-carboxyl group have been corrected from their original X-ray assignments. The other two zinc ligands, His 69 and Glu 72, were identified previously from chemical and X-ray studies. The assignment of the two half-cystinyl residues and the postulation of the existence of a disulfide bond have been confirmed.

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Selected References

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

  1. BARGETZI J. P. KUMAR KS, COX DJ, WALSH KA, NEURATH H: THE AMINO ACID COMPOSITION OF BOVINE PANCREATIC CARBOXYPEPTIDASE A. Biochemistry. 1963 Nov-Dec;2:1468–1474. doi: 10.1021/bi00906a046. [DOI] [PubMed] [Google Scholar]
  2. BARGETZI J. P., THOMPSON E. O., SAMPATHKUMAR K. S., WALSH K. A., NEURATH H. THE AMINO- AND CARBOXYL-TERMINAL RESIDUES AND THE SELF-DIGESTION OF BOVINE PANCREATIC CARBOXYPEPTIDASE A. J Biol Chem. 1964 Nov;239:3767–3774. [PubMed] [Google Scholar]
  3. Bradshaw R. A., Babin D. R., Nomoto M., Srinivasin N. G., Ericsson L. H., Walsh K. A., Neurath H. The amino acid sequence of bovine carboxypeptidase A. II. Tryptic and chymotryptic peptides of the cyanogen bromide fragment F-III. Biochemistry. 1969 Sep;8(9):3859–3871. doi: 10.1021/bi00837a052. [DOI] [PubMed] [Google Scholar]
  4. Bradshaw R. A., Neurath H., Walsh K. A. Considerations of the concept of structural homology as applied to bovine carboxypeptidases A and B. Proc Natl Acad Sci U S A. 1969 Jun;63(2):406–411. doi: 10.1073/pnas.63.2.406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. COX D. J., BOVARD F. C., BARGETZI J. P., WALSH K. A., NEURATH H. PROCEDURES FOR THE ISOLATION OF CRYSTALLINE BOVINE PANCREATIC CARBOXYPEPTIDASE A. II. ISOLATION OF CARBOXYPEPTIDASE A-ALPHA FROM PROCARBOXYPEPTIDASE A. Biochemistry. 1964 Jan;3:44–47. doi: 10.1021/bi00889a008. [DOI] [PubMed] [Google Scholar]
  6. KUMAR K. S., CLEGG J. B., WALSH K. A. THE N-TERMINAL SEQUENCE OF BOVINE CARBOXYPEPTIDASE A AND ITS RELATION OF ZYMOGEN ACTIVATION. Biochemistry. 1964 Nov;3:1728–1732. doi: 10.1021/bi00899a024. [DOI] [PubMed] [Google Scholar]
  7. NEURATH H. MECHANISM OF ZYMOGEN ACTIVATION. Fed Proc. 1964 Jan-Feb;23:1–7. [PubMed] [Google Scholar]
  8. Nomoto M., Srinivasan N. G., Bradshaw R. A., Wade R. D., Neurath H. The amino acid sequence of bovine carboxypeptidase A. I. Preparation and properties of the fragments obtained by cyanogen bromide cleavage. Biochemistry. 1969 Jul;8(7):2755–2762. doi: 10.1021/bi00835a010. [DOI] [PubMed] [Google Scholar]
  9. Nomoto M., Srinivasan N. G., Bradshaw R. A., Wade R. D., Neurath H. The amino acid sequence of bovine carboxypeptidase A. I. Preparation and properties of the fragments obtained by cyanogen bromide cleavage. Biochemistry. 1969 Jul;8(7):2755–2762. doi: 10.1021/bi00835a010. [DOI] [PubMed] [Google Scholar]
  10. Pétra P. H., Bradshaw R. A., Walsh K. A., Neurath H. Identification of the amino acid replacements characterizing the allotypic forms of bovine carboxypeptidase A. Biochemistry. 1969 Jul;8(7):2762–2768. doi: 10.1021/bi00835a011. [DOI] [PubMed] [Google Scholar]
  11. QUIOCHO F. A., RICHARDS F. M. INTERMOLECULAR CROSS LINKING OF A PROTEIN IN THE CRYSTALLINE STATE: CARBOXYPEPTIDASE-A. Proc Natl Acad Sci U S A. 1964 Sep;52:833–839. doi: 10.1073/pnas.52.3.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Riordan J. F., Sokolovsky M., Vallee B. L. The functional tyrosyl residues of carboxypeptidase A. Nitration with tetranitromethane. Biochemistry. 1967 Nov;6(11):3609–3617. doi: 10.1021/bi00863a036. [DOI] [PubMed] [Google Scholar]

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