Crystallization and preliminary X-ray diffraction analysis of selenophosphate synthetases from Trypanosoma brucei and Leishmania major
Crystallization and preliminary X-ray diffraction analysis of selenophosphate synthetases from Trypanosoma brucei and Leishmania major
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DOI：
10.1107/s1744309113014632
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发表时间：
2013-08-01
影响因子：
0.9
通讯作者：
Thiemann, Otavio Henrique
中科院分区：
文献类型：
--
作者：
Faim, Livia Maria;Rosa e Silva, Ivan;Thiemann, Otavio Henrique
硒磷酸合成酶（Selenophosphate synthetase， SPS）在硒代谢中起着不可缺少的作用，它负责催化硒化物与腺苷5′-三磷酸（adenosine 5’-triphosphate， ATP）活化生成硒磷酸，硒磷酸是合成硒半胱氨酸必需的硒供体。重组全长利什曼原虫主要SPS （LmSPS2）难以结晶。因此，利用有限的蛋白水解技术，一个稳定的n端截断结构（Delta N-LmSPS2）产生了合适的晶体。采用石蜡油微批法制备布鲁氏锥虫SPS同源体（TbSPS2）。收集了Delta N-LmSPS2的x射线衍射数据，分辨率为1.9埃，TbSPS2为3.4埃。
Selenophosphate synthetase (SPS) plays an indispensable role in selenium metabolism, being responsible for catalyzing the activation of selenide with adenosine 5'-triphosphate (ATP) to generate selenophosphate, the essential selenium donor for selenocysteine synthesis. Recombinant full-length Leishmania major SPS (LmSPS2) was recalcitrant to crystallization. Therefore, a limited proteolysis technique was used and a stable N-terminal truncated construct (Delta N-LmSPS2) yielded suitable crystals. The Trypanosoma brucei SPS orthologue (TbSPS2) was crystallized by the microbatch method using paraffin oil. X-ray diffraction data were collected to resolutions of 1.9 angstrom for Delta N-LmSPS2 and 3.4 angstrom for TbSPS2.