Characterisation and preliminary X‐ray diffraction analysis of human pancreatic procarboxypeptidase A2

FEBS Letters

Avilés, Francesc X¹  Vendrell, Josep¹  Catasús, Lluis¹  Coll, Miquel²  Garcı́a-Sáez, Isabel²  Reverter, David¹ 

[1]Departament de Bioquı́mica, Facultat de Ciències and Institut de Biologia Fonamental, Universitat Autònoma de Barcelona, 08193 Bellaterra (Barcelona), Spain

[2]Departament de Biologia Molecular i Cellular, Centre d'Investigació i Desenvolupament, CSIC, Jordi Girona 18–26, 08034 Barcelona, Spain

关键词: Carboxypeptidase A2;    Proenzyme;    Substrate specificity;    Crystallography;    proCP;    procarboxypeptidase;    Cbz;    carbobenzoxy;    FAPP;    N-(3-[2-furyl]acryloyl)-l-phenylalanyl-l-phenylalanine;   

DOI  :  10.1016/S0014-5793(97)01476-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 P2₁, and present one procarboxypeptidase A2 molecule per asymmetric unit. The crystals diffract beyond 1.8 Å resolution and are suitable for detailed X-ray analysis.

【 授权许可】

Unknown