Philbrick J B, Zilinskas B A
Department of Biochemistry and Microbiology, Cook College, Rutgers University, New Brunswick, NJ 08903.
Mol Gen Genet. 1988 Jun;212(3):418-25. doi: 10.1007/BF00330845.
Affinity purified, polyclonal antibodies raised against the Photosystem II 33 kDa manganese-stabilizing polypeptide of the spinach oxygen-evolving complex were used to isolate the gene encoding the homologous protein from Synechocystis 6803. Comparison of the amino acid sequence deduced from the Synechocystis psb1 nucleotide sequence with recently published sequences of spinach and pea confirms the homology indicated by antigenic cross-reactivity and shows that the cyanobacterial and higher plant sequences are 43% identical and 63% conserved. Regions of identity, varying in length from 1 to 10 consecutive residues, are distributed throughout the protein. The 28 residues at the amino terminus of the psb1 gene product, characteristic of prokaryotic signal peptides, show homology with the carboxyl-terminal third of the transit sequences of pea and spinach and are most likely needed for the transport of the manganese-stabilizing protein across the thylakoid membrane to its destination of the lumen. Synechocystis mutants which contain a kanamycin resistance gene cassette inserted into the coding region for the 32 kDa polypeptide were constructed. These mutants contain no detectable 32 kDa polypeptide, do not evolve oxygen, and are incapable of photoautotrophic growth.
用针对菠菜放氧复合体光系统II 33 kDa锰稳定多肽制备的亲和纯化多克隆抗体，从集胞藻6803中分离出编码同源蛋白的基因。将从集胞藻psb1核苷酸序列推导的氨基酸序列与最近发表的菠菜和豌豆序列进行比较，证实了抗原交叉反应所表明的同源性，并表明蓝细菌和高等植物序列的同一性为43%，保守性为63%。长度从1到10个连续残基不等的相同区域分布在整个蛋白质中。psb1基因产物氨基末端的28个残基，是原核信号肽的特征，与豌豆和菠菜转运序列羧基末端的三分之一具有同源性，很可能是锰稳定蛋白穿过类囊体膜运输到其内腔目的地所必需的。构建了含有插入到32 kDa多肽编码区的卡那霉素抗性基因盒的集胞藻突变体。这些突变体不含可检测到的32 kDa多肽，不释放氧气，并且不能进行光合自养生长。
