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Molecular cloning, expression, purification and crystallographic analysis of PRRSV 3CL protease.

ABSTRACT: 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.

SUBMITTER: Tian X

PROVIDER: S-EPMC2335157 | biostudies-literature | 2007 Aug

REPOSITORIES: biostudies-literature

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Molecular cloning, expression, purification and crystallographic analysis of PRRSV 3CL protease.

Tian Xinsheng X Feng Youjun Y Zhao Tiezhu T Peng Hao H Yan Jinghua J Qi Jianxun J Jiang Fan F Tian Kegong K Gao Feng F

Acta crystallographica. Section F, Structural biology and crystallization communications 20070728 Pt 8

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, be ...[more]

PMID: 17671377

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