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Title 

Purification, crystallization and X-ray crystallographic analysis of Csm5 in Type III-A Crispr-Cas system

Authors 

Yan AnKwang Hyun ParkMinho LeeWoo-Chan AhnIn-Young BaekT J KimEui-Jeon Woo

Issue Date 

2017

Citation 

BIODESIGN = 한국구조생물학회

Abstract 

The CRISPR-Cas system is a microbial adaptive and heritable immune system, of which mechanism relies on the effector ribonucleoprotein (RNP) complex to degrade foreign genetic elements. The effector surveillance complex of the Type III-A CRISPR-Cas system has five Csm components, Csm1~Csm5. The Csm5 protein is placed in the crRNA 3' ends in the effector RNP complex, leading to speculations that crRNA maturation may be catalyzed by the Csm5 subunit. However, the crystal structure and the detailed function of Csm5 still remain elusive. In this study, the Csm5 from Thermococcus onnurineus NA1 was purified and crystallized by the sitting drop method in the condition of 20% w/v PEG 8000 and 100 mM CHES/sodium hydroxide pH 9.5 at 291K. The diffraction data were collected to a resolution of 3.5 ?. The crystal belonged to space group P4 3 2 1 2, with unit cell parameters a=81.0 ?, b=81.0 ?, c=169.1 ?. One protomer was presented in the asymmetric unit with a corresponding V M of 2.10 ? 3 Da -1 and solvent content of 50%.

ISSN 

2288-6982

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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