嗜热栖热菌HB8中依赖NADP的3-羟基异丁酸脱氢酶的结晶及初步X射线晶体学研究。
Crystallization and preliminary X-ray crystallographic studies of NADP-dependent 3-hydroxyisobutyrate dehydrogenase from Thermus thermophilus HB8.
作者信息
Lokanath Neratur K, Shiromizu Ikuya, Nodake Yuichi, Sugahara Mitsuaki, Yokoyama Shigeyuki, Kuramitsu Seiki, Miyano Masashi, Kunishima Naoki
机构信息
Highthroughput Factory, RIKEN Harima Institute at SPring-8, 1-1-1 Kouto, Mikazuki-cho, Sayo-gun, Hyogo 679-5148, Japan.
出版信息
Acta Crystallogr D Biol Crystallogr. 2003 Dec;59(Pt 12):2294-6. doi: 10.1107/s090744490302002x. Epub 2003 Nov 27.
3-Hydroxyisobutyrate, a central metabolite in the valine catabolic pathway, is reversibly oxidized to methylmalonate semialdehyde by a specific NADP-dependent dehydrogenase (HIBADH). HIBADH from Thermus thermophilus HB8 has been overexpressed in Escherichia coli and crystallized by the microbatch method using lithium chloride as a precipitant at 296 K. X-ray diffraction data have been collected to 1.80 A resolution at 100 K using synchrotron radiation. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 85.878, b = 106.367, c = 168.639 A. A homotetramer of HIBADH is likely to be present in the asymmetric unit, giving a V(M) of 3.0 A(3) Da(-1) and a solvent content of 59.3%.
3-羟基异丁酸是缬氨酸分解代谢途径中的一种中心代谢物，它通过一种特定的依赖于烟酰胺腺嘌呤二核苷酸磷酸（NADP）的脱氢酶（HIBADH）可逆地氧化为甲基丙二酸半醛。嗜热栖热菌HB8的HIBADH已在大肠杆菌中过表达，并通过微量分批法在296 K下使用氯化锂作为沉淀剂进行结晶。利用同步辐射在100 K下收集了分辨率为1.80 Å的X射线衍射数据。晶体属于正交晶系空间群P2(1)2(1)2(1)，晶胞参数a = 85.878，b = 106.367，c = 168.639 Å。非对称单元中可能存在HIBADH的同四聚体，其V(M)为3.0 ų Da⁻¹，溶剂含量为59.3%。
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