Amino acid residues that are critical for in vivo catalytic activity of CtpA, the carboxyl-terminal processing protease for the D1 protein of photosystem II.
CtpA, a carboxyl-terminal processing protease, is a member of a novel family of endoproteases that includes a tail-specific protease from Escherichia coli. In oxygenic photosynthetic organisms, CtpA catalyzes C-terminal processing of the D1 protein of photosystem II, an essential event for the assembly of a manganese cluster and consequent light-mediated water oxidation. We introduced site-specific mutations at 14 conserved residues of CtpA in the cyanobacterium Synechocystis sp. PCC 6803 to examine their functional roles. Analysis of the photoautotrophic growth capabilities of these mutants, their ability to process precursor D1 protein and hence evolve oxygen, along with an estimation of the protease content in the mutants revealed that five of these residues are critical for in vivo activity of CtpA. Recent x-ray crystal structure analysis of CtpA from the eukaryotic alga Scenedesmus obliquus (Liao, D.-I., Qian, J., Chisholm, D. A., Jordan, D. B. and Diner, B. A. (2000) Nat. Struct. Biol. 7, 749-753) has shown that the residues equivalent to Ser-313 and Lys-338, two of the five residues mentioned above, form the catalytic center of this enzyme. Our in vivo analysis demonstrates that the three other residues, Asp-253, Arg-255, and Glu-316, are also important determinants of the catalytic activity of CtpA.
Inagaki
N
N
Department of Biology, Washington University, St. Louis, Missouri 63130-4899, USA.
Maitra
R
R
Satoh
K
K
Pakrasi
H B
HB
D020418:Algal Proteins; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D015153:Blotting, Western; D002268:Carboxypeptidases; D002384:Catalysis; D002455:Cell Division; D017124:Conserved Sequence; D000458:Cyanobacteria / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010100:Oxygen / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D045332:Photosystem II Protein Complex; D043484:Proprotein Convertases; D017386:Sequence Homology, Amino Acid; D012694:Serine / Q000737:chemistry; D013696:Temperature; D013997:Time Factors; D014867:Water / Q000378:metabolism
D020418:Algal Proteins; D000596:Amino Acids; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; C422369:photosystem II, psbA subunit; D014867:Water; D012694:Serine; D002268:Carboxypeptidases; D010450:Endopeptidases; D043484:Proprotein Convertases; C073491:photosystem II, D1 polypeptide carboxy-terminal processing protease; D008239:Lysine; D010100:Oxygen