Pedrotti B, Soffientini A, Islam K
Lepetit Research Center, Marion Merrell-Dow Research Institute, Gerenzano (Va), Italy.
Cell Motil Cytoskeleton. 1993;25(3):234-42. doi: 10.1002/cm.970250304.
The influence of two commonly used sulphonate buffers, PIPES and MES, on the in vitro assembly of bovine brain microtubule protein was examined. Microtubule assembly was monitored by turbimetry and, after centrifugation, the polymerised protein was analysed by SDS-PAGE and western blotting. Assembly in MES when compared with PIPES resulted in a higher recovery of microtubule proteins at both pH 6.4 and pH 6.9 and in an altered protein composition. The buffer pH affected the total amount of protein polymerised but did not significantly affect the protein composition. At both pH conditions the recovery of HMW-MAPs was markedly increased in MES buffer and this increase was mostly due to an increase in the amount of MAP1.
研究了两种常用的磺酸盐缓冲液PIPES和MES对牛脑微管蛋白体外组装的影响。通过比浊法监测微管组装，离心后，通过SDS-PAGE和蛋白质印迹分析聚合的蛋白质。与PIPES相比，在MES中组装时，在pH 6.4和pH 6.9时微管蛋白的回收率更高，且蛋白质组成发生了变化。缓冲液pH影响聚合的蛋白质总量，但对蛋白质组成没有显著影响。在两种pH条件下，MES缓冲液中高分子量微管相关蛋白（HMW-MAPs）的回收率均显著增加，且这种增加主要是由于微管相关蛋白1（MAP1）量的增加。
