Crystallization and preliminary X-ray crystallographic analysis of the catalytic domain of pyrrolysyl-tRNA synthetase from the methanogenic archaeon Methanosarcina mazei.
Crystallization and preliminary X-ray crystallographic analysis of the catalytic domain of pyrrolysyl-tRNA synthetase from the methanogenic archaeon Methanosarcina mazei.
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DOI：
10.1107/s1744309106036700
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发表时间：
2006-10
期刊：
影响因子：
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通讯作者：
T. Yanagisawa;R. Ishii;R. Fukunaga;O. Nureki;S. Yokoyama
中科院分区：
文献类型：
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作者：
T. Yanagisawa;R. Ishii;R. Fukunaga;O. Nureki;S. Yokoyama
在大肠杆菌中高效表达了N端截短型PylRS（c270）的马氏甲烷八叠球菌吡咯赖氨酰-tRNA合成酶（PylRS），纯化后用聚乙二醇作为沉淀剂，通过悬滴气相扩散法进行结晶。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 A，α = β = 90，γ = 120度，并且衍射至1.9 A分辨率。不对称单元含有一个PylRS分子（c270）。硒代蛋氨酸取代的蛋白质晶体的制备，以解决结构的MAD定相方法。
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.