Abstract
The O2-evolution deficient mutant (LF-1) of Scenedesmus obliquus inserts an unprocessed D1 protein into the thylakoid membrane and binds less than half the wild type (WT) level of Mn. LF-1 photosystem II (PS II) membrane fragments lack that part of the high-affinity Mn2+-binding site found in WT membranes which may be associated with histidine residues on the D1 protein (Seibert et al. 1989 Biochim Biophys Acta 974: 185–191). Hsu et al. (1987 Biochim Biophys Acta 890: 89–96) purport that the high-affinity site (characterized by competitive inhibition of DPC-supported DCIP photoreduction by μM concentrations of Mn2+) in Mn-extracted PS II membranes is also the binding site for Mn functional in O2 evolution. Proteases (papain, subtilisin, and carboxypeptidase A) can be used to regenerate the high-affinity Mn2+-binding site in LF-1 PS II membranes but not in thylakoids. Experiments with the histidine modifier, DEPC, suggest that the regenerated high-affinity Mn2+-binding sites produced by either subtilisin or carboxypeptidase A treatments were the same sites observed in WT membranes. However, none of the protease treatments produced LF-1 PS II membranes that could be photoactivated. Reassessment of the processing studies of Taylor et al. (1988 FEBS Lett 237: 229–233) lead us to believe that their procedure also does not result in substantial photoactivation of LF-1 PS II membranes. We conclude that (1) the unprocessed carboxyl end of the D1 protein in LF-1 is located on the lumenal side of the PS II membrane, (2) the unprocessed fragment physically obstructs or perturbs that part of the high-affinity Mn2+-binding site undetectable in LF-1, and (3) the D1 protein must be processed at the time of insertion into the membrane for normal O2-evolution function to result.
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Abbreviations
- Chl:
-
chlorophyll
- DCBQ:
-
2,6-dichloro-1,4-benzoquinone
- DCIP:
-
2,6-dichlorophenol indophenol
- DEPC:
-
diethylpryocarbonate
- DPC:
-
1,5-diphenylcarbazide
- HEPES:
-
4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid
- LDS-PAGE:
-
lithium dodecylsulfate polyacrylamide gel electrophoresis
- LF-1:
-
a low-fluorescent mutant of Scenedesmus obliquus
- MES:
-
4-morpholineethanesulfonic acid
- PS II:
-
photosystem II
- PMSF:
-
phenylmethylsulfonyl fluoride
- RC:
-
photosystem II reaction center
- Tris:
-
tris(hydroxymethyl)aminomethane
- WT:
-
wild type
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Preston, C., Seibert, M. Regeneration of the high-affinity manganese-binding site in the reaction center of an oxygen-evolution deficient mutant of Scenedesmus by protease action. Photosynth Res 22, 101–113 (1989). https://doi.org/10.1007/BF00114770
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DOI: https://doi.org/10.1007/BF00114770