Crowlesmith I, Gamon K, Henning U
Eur J Biochem. 1981 Jan;113(2):375-80. doi: 10.1111/j.1432-1033.1981.tb05076.x.
The OmpA and OmpF proteins are major outer membrane proteins of Escherichia coli K12. Their precursors, the pro-OmpA and pro-OmpF proteins, have been detected in vivo in pulse-labelling experiments carried out with [35S]methionine at 25 degrees C. Wehn the pulse was at 37 degrees C, however, no precursors were detected. The pulse-labelled precursors were processed rapidly and quantitatively into mature protein at 25 degrees C. The apparent half-life of the pro-OmpF protein was estimated to be 30 s, and the pro-OmpA protein may be processed even faster. In short pulses (10 s) the precursors of both proteins were the predominant labelled species, indicating that at 25 degrees C processing does not start until chain elongation of the precursor is almost, if not entirely, complete. When French press lysates of cells pulse-labelled for 10 s were subjected to sucrose gradient centrifugation to separate the inner and outer membranes, both precursors comigrated with the inner membrane.
OmpA蛋白和OmpF蛋白是大肠杆菌K12的主要外膜蛋白。在用[35S]甲硫氨酸于25℃进行的脉冲标记实验中,已在体内检测到它们的前体,即前OmpA蛋白和前OmpF蛋白。然而,当脉冲在37℃进行时,未检测到前体。脉冲标记的前体在25℃下迅速且定量地加工成成熟蛋白。前OmpF蛋白的表观半衰期估计为30秒,前OmpA蛋白的加工速度可能更快。在短脉冲(10秒)中,两种蛋白的前体是主要的标记物种,这表明在25℃下,加工直到前体的链延伸几乎(如果不是完全)完成才开始。当对脉冲标记10秒的细胞进行法国压榨裂解物进行蔗糖梯度离心以分离内膜和外膜时,两种前体都与内膜一起迁移。