Green B R
Biochim Biophys Acta. 1980 Aug 26;609(1):107-20. doi: 10.1016/0005-2787(80)90205-1.
Acetabularia chloroplasts can incorporate radioactive amino acids for up to several hours in vitro. The incorporation is sensitive to chloramphenicol and lincomycin, insensitive to cycloheximide, and completely light-dependent. At least 35 discrete labelled bands can be separated by SDS-polyacrylamide gel electrophoresis: 20--24 in the soluble fraction and 13--15 in the membrane fraction. Most of the label (80--85%) is in the membrane fraction, and 90% of that is in a polypeptide of 32 000 daltons. Chlorophyll-protein complexes were purified from in vitro labelled chloroplasts by SDS-polyacrylamide gel electrophoresis. CP I (P-700-chlorophyll alpha-protein complex) and its apoprotein were both labelled. This shows that the apoprotein is synthesized on chloroplast ribosomes, and can be integrated correctly into the thylakoid membrane in the absence of any cytoplasmic contribution. In contrast, no label was incorporated into the two polypeptides of CP II, the light-harvesting chlorophyll a/b complex.
伞藻叶绿体在体外可摄取放射性氨基酸长达数小时。这种摄取对氯霉素和林可霉素敏感，对环己酰亚胺不敏感，且完全依赖光照。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳可分离出至少35条离散的标记带：20 - 24条在可溶性部分，13 - 15条在膜部分。大部分标记（80 - 85%）在膜部分，其中90%存在于一条32000道尔顿的多肽中。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳从体外标记的叶绿体中纯化叶绿素 - 蛋白质复合物。光系统I（P - 700 - 叶绿素α - 蛋白质复合物）及其脱辅基蛋白均被标记。这表明脱辅基蛋白是在叶绿体核糖体上合成的，并且在没有任何细胞质参与的情况下能够正确整合到类囊体膜中。相比之下，光捕获叶绿素a/b复合物光系统II的两条多肽未被标记。
