Lokanath Neratur K, Shiromizu Ikuya, Nodake Yuichi, Sugahara Mitsuaki, Yokoyama Shigeyuki, Kuramitsu Seiki, Miyano Masashi, Kunishima Naoki
Highthroughput Factory, RIKEN Harima Institute at SPring-8, 1-1-1 Kouto, Mikazuki-cho, Sayo-gun, Hyogo 679-5148, Japan.
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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