Badasso Mohammed O, Dhanaraj V, Wood S P, Cooper J B, Blundell T L
Department of Oral Science, University of Minnesota, Minneapolis, MN 55455, USA.
Acta Crystallogr D Biol Crystallogr. 2004 Apr;60(Pt 4):770-2. doi: 10.1107/S0907444904003221. Epub 2004 Mar 23.
Y75N mutant Mucor pusillus pepsin has been overexpressed in yeast, purified and cocrystallized with the iodine-containing human renin inhibitor CP-113972 [(2R,3S]-isopropyl 3-[(L-prolyl-p-iodo-L-phenylalanyl-S-methyl-cysteinyl)amino-4]-cyclohexyl-2-hydroxybutanoate] for X-ray crystallography. Tetragonal complex crystals with space group P4(3)2(1)2 were produced by the hanging-drop vapour-diffusion method and diffracted to 3.0 A. The crystals exhibited unit-cell parameters a = b = 182.5, c = 99.1 A and contained four molecules in the asymmetric unit. A 96% complete data set was collected at 298 K using Cu Kalpha X-rays from a rotating-anode generator. Solution of the crystal structure of Y75N mutant M. pusillus pepsin is under way by molecular replacement using the molecular coordinates of wild-type M. pusillus pepsin as a model.
Y75N突变型微小毛霉胃蛋白酶已在酵母中过表达、纯化，并与含碘的人肾素抑制剂CP-113972 [（2R,3S]-异丙基3-[(L-脯氨酰-p-碘-L-苯丙氨酰-S-甲基-半胱氨酰)氨基-4]-环己基-2-羟基丁酸酯] 共结晶用于X射线晶体学研究。通过悬滴气相扩散法得到了空间群为P4(3)2(1)2的四方晶系复合晶体，其衍射分辨率达到3.0 Å。这些晶体的晶胞参数a = b = 182.5，c = 99.1 Å，不对称单元中包含四个分子。使用旋转阳极发生器产生的Cu Kα X射线在298 K下收集了96%完整的数据集。目前正在以野生型微小毛霉胃蛋白酶的分子坐标为模型，通过分子置换法解析Y75N突变型微小毛霉胃蛋白酶的晶体结构。
