Abstract
The intrinsic chlorophyll-protein CP 47 is a component of photosystem II which functions in both light-harvesting and oxygen evolution. The large extrinsic loop E of this protein has been shown to interact with the oxygen-evolving site. Previously, Vermaas and coworkers have produced a number of deletions within loop E which yielded mutants which were unable to grow photoautotrophically and which could not evolve oxygen at normal rates. During the course of our site-directed mutagenesis program in Synechocystis 6803, we have altered all of the conserved charged residues which were present within six of these deletions. All ten of these mutants were photoautotrophic and evolved oxygen at normal rates. We speculate that the severe phenotypes of the deletion mutants observed by Vermaas and coworkers in due to large structural perturbations in the extrinsic loop E of CP 47.
This is a preview of subscription content, log in via an institution to check access.
References
Bricker TM, Odom WR, Queirolo CB: Clone association of the 33 kDa extrinsic protein with the apoprotein if CPa-1 in photosystem II. FEBS Lett 231: 111–117 (1988).
Bricker TM: The structure and function of CPa-1 and CPa-2 in photosystem II. Photosyn Res 24: 1–13 (1990).
Bricker TM: Oxygen evolution in the absence of the 33 kDa manganese-stabilizing protein. Biochemistry 31: 623–628 (1992).
Burnap RL, Sherman LA: Deletion mutagenesis in Synechocystis sp. PCC 6803 indicates that the Mn-stabilizing protein of photosystem II is not essential for oxygen evolution. Biochemistry 30: 440–446 (1991).
Debus RJ: The manganese and calcium ions of photosynthetic oxygen evolution. Biochim Biophys Acta 1102: 269–352 (1992).
Eaton-Rye JJ, Vermaas WFJ: Oligonucleotide-directed mutagenesis of psbB, the gene encoding CP 47, employing a deletion strain of the cyanobacterium Synechocystis sp. PCC 6803. Plant Mol Biol 17: 1165–1177 (1991).
Enami I, Kaneko M, Kitamura N, Koike H, Sonoike K, Inoue Y, Katoh S: Total immobilization of the extrinsic 33 kDa protein in spinach photosystem II membrane preparations. Protein stoichiometry and stabilization of oxygen evolution. Biochim Biophys Acta 1060: 224–232 (1991).
Frankel LK, Bricker TM: Mapping of NHS-biotinylation sites and the epitope of the monoclonal antibody FAC2 on the apo-protein of CPa-1. In: Batcheffsky M (ed) Current Research in Photosynthesis, Vol. I, pp. 639–642, Kluwer Academic Publishers, Dordrecht (1990).
Frankel LK, Bricker TM: Interaction of CPa-1 with the manganese-stabilizing protein of photosystem II: Identification of domains on Cpa-1 which are shielded from N-hydroxysuccinimide biotinylation of the manganese-stabilizing protein. Biochemistry 31: 11059–11063 (1992).
Haag E, Eaton-Rye JJ, Renger G and Vermaas WFJ: Functionally important domains of the large hydrophilic loop of CP 47 as probed by oligonucleotide-directed mutagenesis in Synechocytiis sp. PCC 6803. Biochemistry 32: 4444–4454 (1993).
Hayashi H, Fujimura Y, Mohanty PS, Murata N: The role of CP 47 in the evolution of oxygen and the binding of the extrinsic 33-kDa protein to the core complex photosystem II as determined by limited proteolysis. Photosyn Res 36: 35–42 (1993).
Kunkel TA: Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82: 488–499 (1985).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory. Cold Spring Harbor (NY).
Morris A, Herrmann RG: Nucleotide sequence of the gene for the P680 chlorophyll a apoprotein of the photosystem II reaction center from spinach. Nucl Acids Res 12: 2837–2850 (1984).
Odom WR, Bricker TM: Interaction of CPa-1 with the manganese stabilizing protein of photosystem II: Identification of domains crosslinked by 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide. Biochemistry 31: 5616–5620 (1992).
Putnam-Evans C, Bricker TM: Site-directed mutagenesis of the CPa-1 protein of photosystem II: Alteration of the basis resudue pair 384,385R to 384,385G leads to defect associated with the oxygen-evoling complex. Biochemistry 31: 11482–11488 (1992).
Putnam-Evans C, Bricker TM: Site-directed mutagenesis of the CPa-1 protein of photosystem II: Alteraction of the basis residue pair 384,385R to 384,385G leads to a defect associated with the oxygen-evolving complex. Biochemistry 31: 11482–11488 (1992).
Putnam-Evans C, Bricker TM: Site-directed mutagenesis of the CP 47 protein of photosystem II: Alteration of the basic residue 448R to 448G prevents the assembly of functional photosystem II centers under chloride-limiting conditions. Biochemistry 33: 10770–10776 (1994).
Vermaas WFJ, Williams JGK, Arntzen CJ: Sequencing and modification of psbB, the gene encoding the CP 47 protein of photosystem II in the cyanobacterium Synechocystis 6803. Plant Mol Biol 8: 317–326 (1987).
Vermaas WFJ, Ikeuchi M and Inoue Y: Protein composition of the photosystem II core complex in genetically engineered mutants of the cyanobacterium Synechocystis PCC 6803. Photosyn Res 17: 97–113 (1988).
Williams JGK: Construction of specific mutations in photosystem II reaction center by genetic engineering methods in Synechocystis 6803. Meth Enzymol 167: 766–778 (1988).
Putnam-Evans C, Wu J, Burnap R, Whitmarsh J, Bricker TM: Site-directed mutagenesis of the CP 47 protein of photosystem II: alteration of conserved charged residues in the domain 364E-444R. Biochemistry 35: 4046–4053 (1996).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Putnam-Evans, C., Wu, J. & Bricker, T.M. Site-directed mutagenesis of the CP 47 protein of photosystem II: alteration of conserved charged residues which lie within lethal deletions of the large extrinsic loop E. Plant Mol Biol 32, 1191–1195 (1996). https://doi.org/10.1007/BF00041405
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00041405