Crystallization and preliminary X-ray analysis of crustacean hyperglycemic hormone of the kuruma prawn Marsupenaeus japonicus in its weakly-active precursor form
Crystallization and preliminary X-ray analysis of crustacean hyperglycemic hormone of the kuruma prawn Marsupenaeus japonicus in its weakly-active precursor form
复制标题
弱活性前体形式的日本对虾甲壳类高血糖激素的结晶和初步 X 射线分析
DOI：
10.1107/s1744309111040140
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
井上宏隆(永井千晶:第三著者)
中科院分区：
文献类型：
--
作者：
Take Matsuyama;Takahiro Yamashita;Yasushi Imamato;Yoshinori Shichida;Take Matsuyama;井上宏隆(永井千晶:第三著者)
甲壳动物高血糖激素（CHH）在甲壳动物糖代谢的调节中起着关键作用。Pej-SGP-I是日本囊对虾（Marsupenaeus）中已知的六种CHH之一，在大肠杆菌中异源表达，其N-末端带有His标签和Nus标签的蛋白质为弱活性前体形式Pej-SGP-I-Gly，在C-末端有一个额外的甘氨酸残基。以NaCl为主要沉淀剂，采用坐滴法对重组肽进行亲和纯化、标签去除、进一步纯化和结晶。晶体衍射分辨率为1.95 μ m，空间群为原始正交晶系P212121，晶胞参数a = 40.19，B = 53.65，c = 53.63 μ m。 马修斯系数（VM = 1.73 3 Da-1）表明该晶体每个不对称单元含有两个Pej-SGP-I-Gly分子，溶剂含量为29.0%。
Crustacean hyperglycaemic hormone (CHH) plays a pivotal role in the regulation of glucose metabolism in crustaceans. Pej-SGP-I, one of the six known CHHs in the kuruma prawn Marsupenaeus japonicus, was heterologously expressed in Escherichia coli as an N-terminally His-tagged and Nus-tagged protein in its weakly active precursor form, Pej-SGP-I-Gly, which has an extra glycine residue at the C-terminus. The recombinant peptide was subjected to affinity purification, tag removal, further purification and crystallization by the sitting-drop vapour-diffusion method using NaCl as the main precipitant. The crystals diffracted to 1.95 Å resolution and the space group was assigned as primitive orthorhombic P212121, with unit-cell parameters a = 40.19, b = 53.65, c = 53.63 Å. The Matthews coefficient (VM = 1.73 Å3 Da−1) indicated that the crystal contained two Pej-SGP-I-Gly molecules per asymmetric unit, with a solvent content of 29.0%.