Mitchell C, Morris P W, Vary J C
Department of Biochemistry, University of Illinois, Chicago 60612.
J Bacteriol. 1992 Apr;174(8):2474-7. doi: 10.1128/jb.174.8.2474-2477.1992.
Protein phosphorylation in Bacillus subtilis was assayed in vitro by using extracts prepared from cells at various times during growth and sporulation. At least six proteins were labeled in vitro by using [gamma-32P]ATP and extracts of vegetative cells. In extracts prepared at the end of exponential growth and during stationary phase, 12 to 13 proteins were labeled. Seven of the phosphoproteins were purified by fast-performance liquid chromatography and polyacrylamide gel electrophoresis, blotted to Immobilon membranes, and subjected to partial protein sequencing. One of the sequences had sequence homology (greater than 45%) to elongation factor G from several bacterial species, and four sequences matched the predicted amino-terminal sequences of the outB, orfY-tsr, orfU, and ptsH genes.
通过使用在生长和孢子形成的不同时间从细胞制备的提取物，在体外测定枯草芽孢杆菌中的蛋白质磷酸化。使用[γ-32P]ATP和营养细胞提取物，至少有六种蛋白质在体外被标记。在指数生长末期和稳定期制备的提取物中，有12至13种蛋白质被标记。七种磷酸化蛋白通过快速高效液相色谱和聚丙烯酰胺凝胶电泳进行纯化，印迹到Immobilon膜上，并进行部分蛋白质测序。其中一个序列与几种细菌的延伸因子G具有序列同源性（大于45%），并且有四个序列与outB、orfY-tsr,、orfU和ptsH基因的预测氨基末端序列匹配。
