title: Characterisation and preliminary X-ray diffraction analysis of human pancreatic procarboxypeptidase A2.
abstract: Human procarboxypeptidase A2 has been expressed in a Pichia pastoris heterologous system and purified by hydrophobic interaction and anion exchange chromatographies. The hydrolytic action of carboxypeptidase A2 on peptide substrates with different lengths and residues at the C-terminus was analysed, and a preference towards long substrates with aromatic amino acids in their C-terminal end, particularly tryptophan, was found; with such substrates its activity is similar or higher than that of bovine carboxypeptidase A1. Procarboxypeptidase A2 has been crystallised using a vapour diffusion approach; the crystals obtained belong to the monoclinic system, spacegroup P2(1), and present one procarboxypeptidase A2 molecule per asymmetric unit. The crystals diffract beyond 1.8 A resolution and are suitable for detailed X-ray analysis.
authors.affiliation: Department de Bioquímica, Facultat de Ciències and Institut de Biologia Fonamental, Universitat Autònoma de Barcelona, Bellaterra, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Reverter
authors.forename: I
authors.initials: I
authors.lastname: García-Sáez
authors.forename: L
authors.initials: L
authors.lastname: Catasús
authors.forename: J
authors.initials: J
authors.lastname: Vendrell
authors.forename: M
authors.initials: M
authors.lastname: Coll
authors.forename: F X
authors.initials: FX
authors.lastname: Avilés
mesh_terms: D000818:Animals; D002268:Carboxypeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D043422:Carboxypeptidases A; D002417:Cattle; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D018360:Crystallography, X-Ray; D004792:Enzyme Precursors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009842:Oligopeptides / Q000378:metabolism; D010843:Pichia; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D013379:Substrate Specificity
chemical_list: D004792:Enzyme Precursors; D009842:Oligopeptides; D011994:Recombinant Proteins; D002268:Carboxypeptidases; D043422:Carboxypeptidases A