Open AccessPurification, crystallization and preliminary X‐ray analysis of an aminoacylhistidine dipeptidase (PepD) from Vibrio alginolyticus
Author(s) -
Chang ChinYuan,
Hsieh YinCheng,
Wang TingYi,
Chen ChunJung,
Wu TungKung
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910900092x
Subject(s) - dipeptidase , vibrio alginolyticus , enzyme , crystallization , chemistry , dipeptide , monomer , stereochemistry , biochemistry , vibrio , bacteria , biology , amino acid , organic chemistry , polymer , genetics
The aminoacylhistidine dipeptidase (PepD) protein encoded by Vibrio alginolyticus pepD was successfully overexpressed and characterized and the putative active‐site residues responsible for metal binding and catalysis were identified. The purified enzyme contained two zinc ions per monomer. The recombinant dipeptidase enzyme, which was identified as a homodimer in solution, exhibited broad substrate specificity for Xaa‐His dipeptides, with highest activity towards the His‐His dipeptide. The purified protein was crystallized using the hanging‐drop vapour‐diffusion method. Preliminary crystallographic analysis showed that the crystal belonged to space group P 6 1 or P 6 5 , with unit‐cell parameters a = b = 80.42, c = 303.11 Å. The crystal contained two molecules per asymmetric unit and the predicted solvent content was 53.4%.