Lehel C, Oláh Z, Petrovics G, Jakab G, Anderson W B
Laboratory of Cellular Oncology, NCI, NIH, Bethesda, Maryland 20892, USA.
Biochem Biophys Res Commun. 1996 Jun 5;223(1):98-103. doi: 10.1006/bbrc.1996.0852.
Subcellular redistribution (translocation) was initiated by treatment of NIH 3T3 cells overexpressing different epitope-tagged fragments of PKC epsilon with PMA, and was analyzed by immunocytochemistry. The PMA-induced translocation of holo PKC epsilon, as well as fragments epsilon 2 (zinc finger domain + pseudosubstrate domain) and epsilon 7 (zinc finger domain + hinge region) from the Golgi to the plasma membrane was rapid (<10 min), while translocation of fragment epsilon 3 (zinc finger domain) was much slower (30-60 min). These results, combined with results of studies carried out at 20 degrees C to inhibit exocytotic vesicle traffic, indicated that PMA-induced translocation from the Golgi to the plasma membrane may proceed by two distinct mechanisms: a rapid, vesicle independent process noted with holo PKC epsilon (which requires the presence of the pseudosubstrate and/or hinge regions), and a slow, vesicle-dependent pathway observed with the zinc finger fragment.
通过用佛波酯（PMA）处理过表达PKCε不同表位标签片段的NIH 3T3细胞来启动亚细胞重分布（易位），并通过免疫细胞化学进行分析。PMA诱导的全酶PKCε以及片段ε2（锌指结构域+假底物结构域）和ε7（锌指结构域+铰链区）从高尔基体向质膜的易位很快（<10分钟），而片段ε3（锌指结构域）的易位则慢得多（30 - 60分钟）。这些结果，结合在20℃进行的抑制胞吐小泡运输的研究结果，表明PMA诱导的从高尔基体到质膜的易位可能通过两种不同机制进行：一种是全酶PKCε所具有的快速、不依赖小泡的过程（这需要假底物和/或铰链区的存在），另一种是锌指片段所观察到的缓慢、依赖小泡的途径。
