Bhaya D, Jagendorf A T
Arch Biochem Biophys. 1985 Feb 15;237(1):217-23. doi: 10.1016/0003-9861(85)90272-3.
Washed thylakoids of pea chloroplasts, containing tightly bound polysomes, incorporate radioactive amino acids into protein when supplied with soluble factors from Escherichia coli. Polyacrylamide gel electrophoresis with lithium dodecyl sulfate, followed by autoradiography of the labeled products, showed the synthesis of a number of different polypeptides. Two of the most heavily labeled products were in the region expected for the alpha and beta subunits of coupling factor 1, at 57 and 54 kDa. Positive identification of the subunits was made using monospecific antibodies. Furthermore, the same two polypeptides made by soluble polysomes located in the chloroplast stroma were found. While the major proportion of the newly formed alpha and beta subunits made by thylakoid-bound polysomes remained with the thylakoids after protein synthesis occurred, no evidence was found of incorporation into complete, EDTA-extractable coupling factor 1.
豌豆叶绿体的洗涤类囊体含有紧密结合的多核糖体，当提供来自大肠杆菌的可溶性因子时，能将放射性氨基酸掺入蛋白质中。用十二烷基硫酸锂进行聚丙烯酰胺凝胶电泳，随后对标记产物进行放射自显影，结果显示合成了许多不同的多肽。其中两条标记最重的产物位于预期的偶联因子1的α和β亚基所在区域，分子量分别为57 kDa和54 kDa。使用单特异性抗体对亚基进行了阳性鉴定。此外，还发现叶绿体基质中可溶性多核糖体合成了相同的两条多肽。虽然类囊体结合的多核糖体合成的新形成的α和β亚基在蛋白质合成后大部分仍与类囊体结合，但未发现有证据表明它们掺入了完整的、可被EDTA提取的偶联因子1中。
