Vermaas W F, Steinback K E, Arntzen C J
Arch Biochem Biophys. 1984 May 15;231(1):226-32. doi: 10.1016/0003-9861(84)90382-5.
In order to distinguish between two photosystem II proteins with apparent molecular weights of about 32 kDa, mild extraction procedures were used to remove several thylakoid membrane components. A 32-kDa protein that stained intensely with Coomassie brilliant blue could be extracted from the thylakoid membranes without removing the 32-kDa herbicide receptor protein, which stained poorly with Coomassie brilliant blue. The nonextracted protein was readily detectable after in vivo polypeptide labeling with [35S]methionine or after in vitro covalent tagging with [14C]azidoatrazine. The procedures used to extract the intensely stained, 32-kDa polypeptide resulted in changes in herbicide-binding characteristics, presumably due to conformational changes in the herbicide-binding environment. Alterations of membrane surface charge by protein phosphorylation also influenced herbicide binding.
为了区分两种表观分子量约为32 kDa的光系统II蛋白，采用温和的提取程序去除了几种类囊体膜成分。一种用考马斯亮蓝强烈染色的32 kDa蛋白可以从类囊体膜中提取出来，而不会去除用考马斯亮蓝染色较差的32 kDa除草剂受体蛋白。在用[35S]甲硫氨酸进行体内多肽标记后或用[14C]叠氮阿特拉津进行体外共价标记后，未提取的蛋白很容易被检测到。用于提取强烈染色的32 kDa多肽的程序导致除草剂结合特性发生变化，这可能是由于除草剂结合环境的构象变化所致。蛋白质磷酸化引起的膜表面电荷改变也影响除草剂的结合。
