Cloning, crystallization and preliminary X-ray study of XC1258, a CN-hydrolase superfamily protein from Xanthomonas campestris.
CN-hydrolase superfamily proteins are involved in a wide variety of non-peptide carbon-nitrogen hydrolysis reactions, producing some important natural products such as auxin, biotin, precursors of antibiotics etc. These reactions all involve attack on a cyano or carbonyl carbon by a conserved novel catalytic triad Glu-Lys-Cys through a thiol acylenzyme intermediate. However, classification into the CN-hydrolase superfamily based on sequence similarity alone is not straightforward and further structural data are necessary to improve this categorization. Here, the cloning, expression, crystallization and preliminary X-ray analysis of XC1258, a CN-hydrolase superfamily protein from the plant pathogen Xanthomonas campestris (Xcc), are reported. The SeMet-substituted XC1258 crystals diffracted to a resolution of 1.73 A. They are orthorhombic and belong to space group P2(1)2(1)2, with unit-cell parameters a = 143.8, b = 154.63, c = 51.3 A, respectively.
Tsai
Ying-Der
YD
Institute of Biochemistry, National Chung-Hsing University, Taichung 40227, Taiwan.
Chin
Ko-Hsin
KH
Shr
Hui-Lin
HL
Gao
Fei Philip
FP
Lyu
Ping-Chiang
PC
Wang
Andrew H-J
AH
Chou
Shan-Ho
SH
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D017173:Caenorhabditis elegans / Q000235:genetics; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D016959:Xanthomonas campestris / Q000201:enzymology*
D001426:Bacterial Proteins; D006867:Hydrolases