Cupp-Vickery J R, Li H, Poulos T L
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
Proteins. 1994 Oct;20(2):197-201. doi: 10.1002/prot.340200210.
Cytochrome P450eryF was overexpressed in Escherichia coli and purified in high yield. Crystals of the protein in the presence of the substrate, 6-deoxyerythronolide B, have been obtained by the hanging drop vapor diffusion method, using polyethylene glycol 4000 as a precipitant. The crystals belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell dimensions of a = 54.16 A, b = 79.67 A, and c = 99.48 A and one molecule per asymmetric unit. A complete native data set has been collected to a resolution of 2.1 A, and anomalous dispersion difference Patterson maps have revealed the location of the single heme iron atom.
细胞色素P450eryF在大肠杆菌中过表达并以高产率纯化。利用聚乙二醇4000作为沉淀剂，通过悬滴气相扩散法获得了该蛋白在底物6-脱氧红霉内酯B存在下的晶体。这些晶体属于正交晶系空间群P2(1)2(1)2(1)，晶胞参数为a = 54.16 Å，b = 79.67 Å，c = 99.48 Å，每个不对称单元中有一个分子。已收集到分辨率为2.1 Å的完整天然数据集，异常色散差值帕特森图揭示了单个血红素铁原子的位置。
