Münch S, Ljungberg U, Steppuhn J, Schneiderbauer A, Nechushtai R, Beyreuther K, Herrmann R G
Botanisches Institut der Ludwig-Maximilians-Universität München, Federal Republic of Germany.
Curr Genet. 1988 Nov;14(5):511-8. doi: 10.1007/BF00521277.
Several cDNA clones encoding the complete subunit II and III precursor polypeptides of the photosystem I reaction center were isolated from two spinach lambda gt1 1 expression libraries by immunoscreening and homologous hybridization. The identity of the recombinant cDNAs was verified by an N-terminal amino acid sequence of 14 and 20 residues for the respective mature spinach proteins. The ca. 880 nucleotide long sequence and derived amino acid sequence for subunit II predict a precursor of 23.2 kDa (212 residues) and a positively charged, mature protein of 17.9 kDa (162 residues). The corresponding data for subunit III are ca. 710 nucleotides (cDNA), 13.4 kDa (125 residues, precursor polypeptide) and, again, a positively charged, mature protein of 9.7 kDa (91 residues). Secondary structure predictions indicate that both subunits are extramembraneous components of photosystem I. Subunit II is probably located on the matrix-side, subunit III in the lumen of stroma lamellae which is consistent both with biochemical findings and the proposed roles of these proteins in the electron transition from and to photosystem I, respectively. Major transcripts of 1.1 kb (subunit II) and 0.8 kb (subunit III) have been observed by RNA-DNA hybridization.
通过免疫筛选和同源杂交,从两个菠菜λgt11表达文库中分离出了几个编码光系统I反应中心完整亚基II和III前体多肽的cDNA克隆。通过对各自成熟菠菜蛋白14个和20个残基的N端氨基酸序列验证了重组cDNA的身份。亚基II约880个核苷酸长的序列及其推导的氨基酸序列预测其前体为23.2 kDa(212个残基),成熟蛋白带正电荷,为17.9 kDa(162个残基)。亚基III的相应数据约为710个核苷酸(cDNA)、13.4 kDa(125个残基,前体多肽),同样,成熟蛋白带正电荷,为9.7 kDa(91个残基)。二级结构预测表明,这两个亚基都是光系统I的膜外成分。亚基II可能位于基质侧,亚基III位于基质类囊体腔中,这与生化研究结果以及这些蛋白分别在光系统I的电子传递中的作用一致。通过RNA-DNA杂交观察到了1.1 kb(亚基II)和0.8 kb(亚基III)的主要转录本。