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首页> 外文期刊>Acta crystallographica, Section F. Structural biology and crystallization communications >Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human histidine triad nucleotide-binding protein 2 (hHINT2)
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Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human histidine triad nucleotide-binding protein 2 (hHINT2)

机译：人组氨酸三联体核苷酸结合蛋白2（hHINT2）的表达，纯化，结晶和初步X射线晶体学分析

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摘要

Histidine triad nucleotide-binding protein 2 (HINT2) is a mitochondrial adenosine phosphoramidase mainly expressed in the pancreas, liver and adrenal gland. HINT2 possibly plays a role in apoptosis, as well as being involved in steroid biosynthesis, hepatic lipid metabolism and regulation of hepatic mitochondria function. The expression level of HINT2 is significantly down-regulated in hepatocellular carcinoma patients. To date, endogenous substrates for this enzyme, as well as the three-dimensional structure of human HINT2, are unknown. In this study, human HINT2 was cloned, overexpressed in Escherichia coli and purified. Crystallization was performed at 278 K using PEG 4000 as the main precipitant; the crystals, which belonged to the tetragonal space group P4(1)2(1)2 with unit-cell parameters a = b = 76.38, c = 133.25 A, diffracted to 2.83 angstrom resolution. Assuming two molecules in the asymmetric unit, the Matthews coefficient and the solvent content were calculated to be 2.63 angstrom(3) Da(-1) and 53.27%, respectively.
机译：组氨酸三联体核苷酸结合蛋白2（HINT2）是一种线粒体腺苷磷酸酰胺酶，主要表达于胰腺，肝脏和肾上腺。 HINT2可能在细胞凋亡中起作用，并参与类固醇的生物合成，肝脂质代谢和肝线粒体功能的调节。 HINT2的表达水平在肝细胞癌患者中显着下调。迄今为止，该酶的内源底物以及人HINT2的三维结构尚不清楚。在这项研究中，人类HINT2被克隆，在大肠杆菌中过表达并纯化。使用PEG 4000作为主要沉淀剂，在278 K下进行结晶。属于四边形空间群P4（1）2（1）2的晶体，其晶胞参数a = b = 76.38，c = 133.25 A，衍射至2.83埃分辨率。假设两个分子在不对称单元中，马修斯系数和溶剂含量经计算分别为2.63埃（3）Da（-1）和53.27％。

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