人烯醇化磷酸酶E1的纯化、结晶及初步X射线衍射分析
Purification, crystallization and preliminary X-ray diffraction analysis of human enolase-phosphatase E1.
作者信息
Wang Hui, Pang Hai, Ding Yi, Li Yi, Wu Xiao'ai, Rao Zihe
机构信息
Laboratory of Structural Biology, Tsinghua University, Beijing 100084, People's Republic of China.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 May 1;61(Pt 5):521-3. doi: 10.1107/S174430910501184X. Epub 2005 Apr 28.
Enolase-phosphatase E1 (MASA) is a bifunctional enzyme in the ubiquitous methionine-salvage pathway and catalyzes the continuous reaction of 2,3-diketo-5-methylthio-1-phosphopentane to yield the acireductone metabolite. Recombinant human E1 enzyme has been crystallized using the hanging-drop vapour-diffusion method and diffraction-quality crystals were grown at 291 K using PEG 4000 as precipitant. Diffraction data were collected to 1.7 A resolution from SeMet-derivative crystals at 100 K using synchrotron radiation. The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 54.02, b = 57.55, c = 87.32 A. The structure was subsequently solved by the multi-wavelength anomalous diffraction (MAD) phasing method.
烯醇酶磷酸酶E1（MASA）是普遍存在的甲硫氨酸补救途径中的一种双功能酶，催化2,3-二酮-5-甲硫基-1-磷酸戊烷的连续反应以产生酸还原酮代谢物。已使用悬滴气相扩散法对重组人E1酶进行了结晶，并使用聚乙二醇4000作为沉淀剂在291 K下生长出了具有衍射质量的晶体。在100 K下使用同步辐射从SeMet衍生物晶体收集了分辨率为1.7 Å的衍射数据。晶体属于空间群P2(1)2(1)2(1)，晶胞参数a = 54.02，b = 57.55，c = 87.32 Å。随后通过多波长反常衍射（MAD）相位法解析了其结构。
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