White E L, Ross L J, Reynolds R C, Seitz L E, Moore G D, Borhani D W
Drug Discovery Division, Southern Research Institute, Birmingham, AL 35205, USA.
J Bacteriol. 2000 Jul;182(14):4028-34. doi: 10.1128/JB.182.14.4028-4034.2000.
The essential cell division protein, FtsZ, from Mycobacterium tuberculosis has been expressed in Escherichia coli and purified. The recombinant protein has GTPase activity typical of tubulin and other FtsZs. FtsZ polymerization was studied using 90 degrees light scattering. The mycobacterial protein reaches maximum polymerization much more slowly ( approximately 10 min) than E. coli FtsZ. Depolymerization also occurs slowly, taking 1 h or longer under most conditions. Polymerization requires both Mg(2+) and GTP. The minimum concentration of FtsZ needed for polymerization is 3 microM. Electron microscopy shows that polymerized M. tuberculosis FtsZ consists of strands that associate to form ordered aggregates of parallel protofilaments. Ethyl 6-amino-2, 3-dihydro-4-phenyl-1H-pyrido[4,3-b][1,4]diazepin-8-ylcarbamate+ ++ (SRI 7614), an inhibitor of tubulin polymerization synthesized at Southern Research Institute, inhibits M. tuberculosis FtsZ polymerization, inhibits GTP hydrolysis, and reduces the number and sizes of FtsZ polymers.
来自结核分枝杆菌的关键细胞分裂蛋白FtsZ已在大肠杆菌中表达并纯化。该重组蛋白具有微管蛋白和其他FtsZ典型的GTP酶活性。使用90度光散射研究FtsZ聚合。与大肠杆菌FtsZ相比，分枝杆菌蛋白达到最大聚合的速度要慢得多（约10分钟）。解聚也很缓慢，在大多数条件下需要1小时或更长时间。聚合需要Mg(2+)和GTP。聚合所需的FtsZ最低浓度为3 microM。电子显微镜显示，聚合的结核分枝杆菌FtsZ由链组成，这些链相互关联形成平行原丝的有序聚集体。由南方研究所合成的微管蛋白聚合抑制剂6-氨基-2,3-二氢-4-苯基-1H-吡啶并[4,3-b][1,4]二氮杂卓-8-基氨基甲酸乙酯（SRI 7614）可抑制结核分枝杆菌FtsZ聚合，抑制GTP水解，并减少FtsZ聚合物的数量和大小。
