Majewski C, Trebst A
Lehrstuhl Biochemie der Pflanzen, Ruhr-Universität, Bochum, FRG.
Mol Gen Genet. 1990 Dec;224(3):373-82. doi: 10.1007/BF00262431.
A cytochrome bc1-complex of Rs. rubrum was isolated and the three subunits were purified to homogeneity. The N-terminal amino acid sequence of the purified subunits was determined by automatic Edman degradation. The pet genes of Rhodospirillum rubrum coding for the three subunits of the cytochrome bc1-complex were isolated from a genomic library of Rs. rubrum using oligonucleotides specific for conserved regions of the subunits from other organisms and a heterologous probe derived from the genes for the complex of Rb. capsulatus. The complete nucleotide sequence of a 5500 bp SalI/SphI fragment is described which includes the pet genes and three additional unidentified open reading frames. The N-terminal amino acid sequence of the isolated subunits was used for the identification of the three genes. The genes encoding the subunits are organized as follows: Rieske protein, cytochrome b, cytochrome c1. Comparison of the N-terminal protein sequences with the protein sequences deduced from the nucleotide sequence showed that only cytochrome c1 is processed during transport and assembly of the three subunits of the complex. Only the N-terminal methionine of the Rieske protein is cleaved off. The similarity of the deduced amino acid sequence of the three subunits to the corresponding subunits of other organisms is described and implications for structural features of the subunits are discussed.
从深红螺菌中分离出细胞色素bc1复合物,并将三个亚基纯化至同质。通过自动Edman降解法测定纯化亚基的N端氨基酸序列。利用针对其他生物体亚基保守区域的寡核苷酸以及来自荚膜红细菌复合物基因的异源探针,从深红螺菌的基因组文库中分离出编码细胞色素bc1复合物三个亚基的深红螺菌pet基因。描述了一个5500 bp的SalI/SphI片段的完整核苷酸序列,其中包括pet基因和另外三个未鉴定的开放阅读框。分离出的亚基的N端氨基酸序列用于鉴定这三个基因。编码这些亚基的基因组织如下: Rieske蛋白、细胞色素b、细胞色素c1。将N端蛋白质序列与从核苷酸序列推导的蛋白质序列进行比较,结果表明,在复合物的三个亚基的转运和组装过程中,只有细胞色素c1被加工。只有Rieske蛋白的N端甲硫氨酸被切除。描述了三个亚基推导的氨基酸序列与其他生物体相应亚基的相似性,并讨论了其对亚基结构特征的影响。