Lian Wei, Gu Yunrong, Pedersen Brenda, Kukar Thomas, Govindasamy Lakshmanan, Agbandje-McKenna Mavis, Jin Shouguang, McKenna Robert, Wu Donghai
Department of Medicinal Chemistry, McKnight Brain Institute and University of Florida, Gainesville, Florida 32610, USA.
Acta Crystallogr D Biol Crystallogr. 2004 Feb;60(Pt 2):374-5. doi: 10.1107/S090744490302818X. Epub 2004 Jan 23.
Choline acetyltransferase (ChAT) catalyzes the biosynthesis of the neurotransmitter acetylcholine from acetyl-CoA and choline in cholinergic neurons. Rat ChAT (rChAT) was overexpressed in Escherichia coli, purified by affinity chromatography and crystallized. Diffraction data were collected from a single crystal under cryoconditions at the F1 beamline at the Cornell High Energy Synchrotron Source, with a maximal useful diffraction pattern to 1.55 A resolution. The crystals were shown to belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 138.97, b = 77.67, c = 59.67 A and a scaling R(sym) of 0.054 for 72 446 unique reflections. Packing considerations indicate there to be one molecule per asymmetric unit. It is expected that in the near future the structure of rChAT will be obtained using molecular-replacement methods. Elucidation of the structure of rChAT will aid in the development of therapeutic agents for Alzheimer's disease.
胆碱乙酰转移酶（ChAT）在胆碱能神经元中催化由乙酰辅酶A和胆碱合成神经递质乙酰胆碱。大鼠胆碱乙酰转移酶（rChAT）在大肠杆菌中过表达，通过亲和层析纯化并结晶。在康奈尔高能同步辐射源的F1光束线低温条件下，从单晶收集衍射数据，最大有用衍射图谱分辨率达到1.55 Å。晶体属于正交晶系空间群P2(1)2(1)2(1)，晶胞参数a = 138.97，b = 77.67，c = 59.67 Å，72446个独立反射的比例因子R(sym)为0.054。堆积分析表明每个不对称单元有一个分子。预计在不久的将来，将使用分子置换方法获得rChAT的结构。阐明rChAT的结构将有助于开发治疗阿尔茨海默病的药物。
