Michaels A, Herrin D
Department of Biology, Ben Gurion University of the Negev, Beer Sheva, Israel.
Plant Physiol. 1989 Jan;89(1):100-3. doi: 10.1104/pp.89.1.100.
Cell-free translation of Chlamydomonas reinhardtii RNA in the presence of photosynthetic membranes resulted in association of the herbicide binding (Qb) protein with membranes. Incubation of recovered membranes with high salt did not extract the polypeptide from membranes. Tryptic digestion of in vivo labeled membranes or membranes recovered from in vitro translation mixtures showed that Qb had similar orientation. In vitro translation in the presence of chloroplast membranes from cells exposed to high light intensity restored the membrane associated kinase activity lost by photoinhibition. Thus, in vitro synthesis resulted in functional integration of the Qb protein within the photosynthetic membrane.
在光合膜存在的情况下对莱茵衣藻RNA进行无细胞翻译,导致除草剂结合(Qb)蛋白与膜结合。用高盐孵育回收的膜并没有从膜中提取出该多肽。对体内标记的膜或从体外翻译混合物中回收的膜进行胰蛋白酶消化表明,Qb具有相似的方向。在高光强度下暴露的细胞的叶绿体膜存在的情况下进行体外翻译,恢复了因光抑制而丧失的膜相关激酶活性。因此,体外合成导致Qb蛋白在光合膜内进行功能整合。