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来自酿酒酵母的甘油单酯脂肪酶Yju3p可溶性变体的纯化、结晶及初步X射线衍射分析

Purification, crystallization and preliminary X-ray diffraction analysis of a soluble variant of the monoglyceride lipase Yju3p from the yeast Saccharomyces cerevisiae.

作者信息

Rengachari Srinivasan, Aschauer Philipp, Sturm Christian, Oberer Monika

机构信息

Institute of Molecular Biology, University of Graz, Humboldtstrasse 50/3, 8010 Graz, Austria.

出版信息

Acta Crystallogr F Struct Biol Commun. 2015 Feb;71(Pt 2):243-6. doi: 10.1107/S2053230X15001557. Epub 2015 Jan 28.

DOI:10.1107/S2053230X15001557
PMID:25664804
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4321484/
Abstract

The protein Yju3p is the orthologue of monoglyceride lipases in the yeast Saccharomyces cerevisiae. A soluble variant of this lipase termed s-Yju3p (38.3 kDa) was generated and purified to homogeneity by affinity and size-exclusion chromatography. s-Yju3p was crystallized in a vapour-diffusion setup at 293 K and a complete data set was collected to 2.4 Å resolution. The crystal form was orthorhombic (space group P212121), with unit-cell parameters a = 77.2, b = 108.6, c = 167.7 Å. The asymmetric unit contained four molecules with a solvent content of 46.4%.

摘要

蛋白质Yju3p是酿酒酵母中甘油单酯脂肪酶的同源物。通过亲和色谱和尺寸排阻色谱法产生并纯化了这种脂肪酶的一种可溶性变体，称为s-Yju3p（38.3 kDa），使其达到均一性。s-Yju3p在293 K的气相扩散装置中结晶，并收集了分辨率为2.4 Å的完整数据集。晶体形式为正交晶系（空间群P212121），晶胞参数a = 77.2、b = 108.6、c = 167.7 Å。不对称单元包含四个分子，溶剂含量为46.4%。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a096/4321484/378035a532ed/f-71-00243-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a096/4321484/5d3e1fbf2e33/f-71-00243-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a096/4321484/807ce6736e56/f-71-00243-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a096/4321484/378035a532ed/f-71-00243-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a096/4321484/5d3e1fbf2e33/f-71-00243-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a096/4321484/807ce6736e56/f-71-00243-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a096/4321484/378035a532ed/f-71-00243-fig3.jpg

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