来自产甲烷古菌马氏甲烷八叠球菌的吡咯赖氨酸-tRNA合成酶催化结构域的结晶及初步X射线晶体学分析。
Crystallization and preliminary X-ray crystallographic analysis of the catalytic domain of pyrrolysyl-tRNA synthetase from the methanogenic archaeon Methanosarcina mazei.
作者信息
Yanagisawa Tatsuo, Ishii Ryohei, Fukunaga Ryuya, Nureki Osamu, Yokoyama Shigeyuki
机构信息
RIKEN Genomic Sciences Center, Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045, Japan.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Oct 1;62(Pt 10):1031-3. doi: 10.1107/S1744309106036700. Epub 2006 Sep 30.
Abstract
Pyrrolysyl-tRNA synthetase (PylRS) from Methanosarcina mazei was overexpressed in an N-terminally truncated form PylRS(c270) in Escherichia coli, purified to homogeneity and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The native PylRS(c270) crystals in complex with an ATP analogue belonged to space group P6(4), with unit-cell parameters a = b = 104.88, c = 70.43 A, alpha = beta = 90, gamma = 120 degrees , and diffracted to 1.9 A resolution. The asymmetric unit contains one molecule of PylRS(c270). Selenomethionine-substituted protein crystals were prepared in order to solve the structure by the MAD phasing method.
摘要
来自马氏甲烷八叠球菌的吡咯赖氨酰 - tRNA合成酶（PylRS）以N端截短形式PylRS(c270)在大肠杆菌中过表达，纯化至同质，并使用聚乙二醇作为沉淀剂通过悬滴气相扩散法结晶。与ATP类似物复合的天然PylRS(c270)晶体属于空间群P6(4)，晶胞参数a = b = 104.88，c = 70.43 Å，α = β = 90，γ = 120°，衍射分辨率达到1.9 Å。不对称单元包含一个PylRS(c270)分子。制备了硒代甲硫氨酸取代的蛋白质晶体，以便通过MAD相位法解析结构。
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