木瓜蛋白酶热稳定变体前体蛋白的结晶及初步X射线衍射研究
Crystallization and preliminary X-ray diffraction studies of the precursor protein of a thermostable variant of papain.
作者信息
Roy Sumana, Choudhury Debi, Chakrabarti Chandana, Biswas Sampa, Dattagupta J K
机构信息
Crystallography and Molecular Biology Division, Saha Institute of Nuclear Physics, 1/AF Bidhannagar, Kolkata 700 064, India.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 May 1;67(Pt 5):634-6. doi: 10.1107/S1744309111010888. Epub 2011 Apr 28.
Abstract
The crystallization of a recombinant thermostable variant of pro-papain has been carried out. The mutant pro-enzyme was expressed in Escherichia coli as inclusion bodies, refolded, purified and crystallized. The crystals belonged to space group P2(1), with unit-cell parameters a = 42.9, b = 74.8, c = 116.5 Å, β = 93.0°, and diffracted to 2.6 Å resolution using synchrotron radiation. Assuming the presence of two molecules in the asymmetric unit, the calculated Matthews coefficient is 2.28 Å(3) Da(-1), corresponding to a solvent content of 46%. Initial attempts to solve the structure using molecular-replacement techniques were successful.
摘要
已完成一种重组热稳定木瓜蛋白酶原变体的结晶。该突变体酶原在大肠杆菌中表达为包涵体，经复性、纯化和结晶。晶体属于空间群P2(1)，晶胞参数a = 42.9、b = 74.8、c = 116.5 Å，β = 93.0°，使用同步辐射衍射至2.6 Å分辨率。假设不对称单位中有两个分子，计算得到的马修斯系数为2.28 Å(3) Da(-1)，对应溶剂含量为46%。最初使用分子置换技术解析结构的尝试取得了成功。
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