Nucleotide sequences of cDNAs encoding the entire precursor polypeptides for subunits II and III of the photosystem I reaction center from spinach

Abstract

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.