Prince S M, Feron C, Janssens D, Lobet Y, Achtman M, Kusecek B, Bullough P A, Derrick J P
Department of Biomolecular Sciences, UMIST, PO Box 88, Manchester, England.
Acta Crystallogr D Biol Crystallogr. 2001 Aug;57(Pt 8):1164-6. doi: 10.1107/s0907444901009416. Epub 2001 Jul 23.
OpcA is an integral outer membrane from the Gram-negative pathogen Neisseria meningitidis that plays a role in adhesion of meningococci to host cells. The protein was overexpressed in Escherichia coli in an insoluble form and a procedure developed for refolding by rapid dilution from denaturant into detergent solution. The refolded material was identical to native OpcA isolated from meningococci, as judged by overall molecular weight, migration on SDS-PAGE and reaction against monoclonal antibodies. Both native and recombinant OpcA crystallized under similar conditions to give an orthorhombic crystal form (P2(1)2(1)2), with unit-cell parameters a = 96.9, b = 46.3, c = 74.0 A. Complete data sets of reflections were collected from native and refolded OpcA to 2.0 A resolution.
OpcA是革兰氏阴性病原体脑膜炎奈瑟菌的一种整合外膜蛋白，在脑膜炎球菌与宿主细胞的黏附中发挥作用。该蛋白在大肠杆菌中以不溶性形式过表达，并开发了一种从变性剂快速稀释到去污剂溶液中进行重折叠的方法。通过整体分子量、SDS-PAGE迁移和针对单克隆抗体的反应判断，重折叠后的物质与从脑膜炎球菌中分离出的天然OpcA相同。天然和重组OpcA在相似条件下结晶，形成正交晶型（P2(1)2(1)2），晶胞参数a = 96.9，b = 46.3，c = 74.0 Å。从天然和重折叠的OpcA收集了完整的反射数据集，分辨率达到2.0 Å。
