Chua N H, Gillham N W
J Cell Biol. 1977 Aug;74(2):441-52. doi: 10.1083/jcb.74.2.441.
The sites of synthesis of the major thylakoid membrane polypeptides have been studied in the green alga Chlamydomonas reinhardtii by pulse labeling of cells with [14C]acetate in the presence of inhibitors specific for chloroplast and cytoplasmic protein synthesis. The labeled membrane polypeptides were separated by an improved method of sodium dodecyl sulfate (SDS) gradient gel electrophoresis, and autoradiographs were made of the dried gels. The results demonstrate that of the 33 polypeptides resolved in the gels, at least nine are made on chloroplast ribosomes. Two of these (polypeptides 2 and 6) are associated with the reaction centers of photosystems I and II. Another polypeptide (polypeptide 5) appears from genetic data to be coded by chloroplast DNA. Experiments with a mutant whose chloroplast ribosomes are resistant to spectinomycyn (spr-u-1-6-2) show that polypeptides whose synthesis takes place on chloroplast ribosomes are made in the presence of spectinomycin in the mutant although their synthesis is blocked by this antibiotic in wild type cells.
通过在存在叶绿体和细胞质蛋白质合成特异性抑制剂的情况下用[14C]乙酸盐对莱茵衣藻细胞进行脉冲标记,研究了主要类囊体膜多肽的合成位点。用改进的十二烷基硫酸钠(SDS)梯度凝胶电泳方法分离标记的膜多肽,并对干燥的凝胶进行放射自显影。结果表明,在凝胶中分辨出的33种多肽中,至少有9种是在叶绿体核糖体上合成的。其中两种(多肽2和6)与光系统I和II的反应中心相关。从遗传数据来看,另一种多肽(多肽5)由叶绿体DNA编码。用一种叶绿体核糖体对壮观霉素具有抗性的突变体(spr-u-1-6-2)进行的实验表明,在突变体中,那些在叶绿体核糖体上合成的多肽是在有壮观霉素存在的情况下合成的,尽管在野生型细胞中这种抗生素会阻断它们的合成。