Molecular cloning, expression, purification and crystallographic analysis of PRRSV 3CL protease.
3CL protease, a viral chymotrypsin-like proteolytic enzyme, plays a pivotal role in the transcription and replication machinery of many RNA viruses, including porcine reproductive and respiratory syndrome virus (PRRSV). In this study, the full-length 3CL protease from PRRSV was cloned and overexpressed in Escherichia coli. Crystallization experiments yielded crystals that diffracted to 2.1 A resolution and belong to space group C2, with unit-cell parameters a = 112.31, b = 48.34, c = 42.88 A, beta = 109.83 degrees . The Matthews coefficient and the solvent content were calculated to be 2.49 A(3) Da(-1) and 50.61%, respectively, for one molecule in the asymmetric unit.
Center for Molecular Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
Xinsheng
X
Tian
Youjun
Y
Feng
Tiezhu
T
Zhao
Hao
H
Peng
Jinghua
J
Yan
Jianxun
J
Qi
Fan
F
Jiang
Kegong
K
Tian
Feng
F
Gao
D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D010447:Peptide Hydrolases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D019316:Porcine respiratory and reproductive syndrome virus / Q000201:enzymology* / Q000235:genetics
D010447:Peptide Hydrolases