title: Structural evolution of an enzyme specificity. The structure of rat carboxypeptidase A2 at 1.9-A resolution.
abstract: The structure of rat carboxypeptidase A2 (CPA2), which has a unique specificity for tryptophan-containing COOH-terminal peptides, has been determined in an unliganded state at 1.9-A resolution and refined to a crystallographic R-factor of 18.3%. Comparison of the structure of CPA2 with that of bovine carboxypeptidase A (referred to here as CPA1) reveals that the specificity of the former for larger amino acids probably arises from two amino acid replacements within the binding cavity (Thr268----Ala and Leu203----Met), coupled with differences in the positions of conserved residues in a surface loop on one face of the specificity pocket. The position of the reactive-site surface loop may be affected also by a disulfide bridge between Cys210 and Cys244. In this unliganded form of the enzyme, Tyr248 takes up a position interior to the specificity pocket and is distinct from that observed in bovine CPA1. The structural differences between CPA1 and CPA2 correlate strongly with crystallographically determined temperature factors and thus appear to be largest where the enzyme is flexible.
authors.affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas 75235-9050.
authors.forename: Z
authors.initials: Z
authors.lastname: Faming
authors.forename: B
authors.initials: B
authors.lastname: Kobe
authors.forename: C B
authors.initials: CB
authors.lastname: Stewart
authors.forename: W J
authors.initials: WJ
authors.lastname: Rutter
authors.forename: E J
authors.initials: EJ
authors.lastname: Goldsmith
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D005075:Biological Evolution; D002268:Carboxypeptidases / Q000737:chemistry* / Q000235:genetics; D043422:Carboxypeptidases A; D002417:Cattle; D007527:Isoenzymes / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D011487:Protein Conformation; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014364:Tryptophan; D014961:X-Ray Diffraction
chemical_list: D007527:Isoenzymes; D013438:Sulfhydryl Compounds; D014364:Tryptophan; D002268:Carboxypeptidases; D043422:Carboxypeptidases A