Nasir Nazia, Vyas Rajan, Biswal Bichitra K
Protein Crystallography Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110 067, India.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Apr 1;69(Pt 4):445-8. doi: 10.1107/S1744309113006210. Epub 2013 Mar 28.
Histidinolphosphate aminotransferase (HisC; Rv1600) from Mycobacterium tuberculosis was overexpressed in M. smegmatis and purified to homogeneity using nickel-nitrilotriacetic acid metal-affinity and gel-filtration chromatography. Diffraction-quality crystals suitable for X-ray analysis were grown by the hanging-drop vapour-diffusion technique using 30% polyethylene glycol monomethyl ether 2000 as the precipitant. The crystals belonged to the hexagonal space group P3221, with an unusual high solvent content of 74.5%. X-ray diffraction data were recorded to 3.08 Å resolution from a single crystal using in-house Cu Kα radiation. The structure of HisC was solved by the molecular-replacement method using its Corynebacterium glutamicum counterpart as a search model. HisC is a dimer in the crystal as well as in solution.
结核分枝杆菌的组氨酸磷酸转氨酶（HisC；Rv1600）在耻垢分枝杆菌中过表达，并通过镍-次氮基三乙酸金属亲和色谱和凝胶过滤色谱纯化至均一。使用30%聚乙二醇单甲醚2000作为沉淀剂，通过悬滴气相扩散技术生长出适合X射线分析的衍射质量晶体。这些晶体属于六方空间群P3221，具有74.5%的异常高溶剂含量。使用内部Cu Kα辐射从单个晶体记录了分辨率为3.08 Å的X射线衍射数据。HisC的结构通过分子置换法解析，使用谷氨酸棒杆菌的对应物作为搜索模型。HisC在晶体和溶液中均为二聚体。
