Purification, crystallization and preliminary X-ray analysis of an aminoacylhistidine dipeptidase (PepD) from Vibrio alginolyticus.
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 P6(1) or P6(5), with unit-cell parameters a = b = 80.42, c = 303.11 A. The crystal contained two molecules per asymmetric unit and the predicted solvent content was 53.4%.
Chang
Chin Yuan
CY
Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan.
Hsieh
Yin Cheng
YC
Wang
Ting Yi
TY
Chen
Chun Jung
CJ
Wu
Tung Kung
TK
D003460:Crystallization; D018360:Crystallography, X-Ray; D004150:Dipeptidases / Q000737:chemistry* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D044149:Vibrio alginolyticus / Q000201:enzymology*
D004150:Dipeptidases; C038937:dipeptidase