Abstract
A method is described for the isolation and purification of active oxygen-evolving photosystem II (PS II) membranes from the green alga Chlamydomonas reinhardtii. The isolation procedure is a modification of methods evolved for spinach (Berthold et al. 1981). The purity and integrity of the PS II preparations have been assesssed on the bases of the polypeptide pattern in SDS-PAGE, the rate of oxygen evolution, the EPR multiline signal of the S2 state, the room temperature chlorophyll a fluorescence yield, the 77 K emission spectra, and the P700 EPR signal at 300 K. These data show that the PS II characteristics are increased by a factor of two in PS II preparations as compared to thylakoid samples, and the PS I concentration is reduced by approximately a factor ten compared to that in thylakoids.
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Abbreviations
- BSA:
-
bovine serum albumin
- Chl:
-
chlorophyll
- DCBQ:
-
2,6-dichloro-p-benzoquinone
- DCMU:
-
(diuron) 3-(3,4-dichlorophenyl)-1,1-dimethylurea
- DMQ:
-
2,5-dimethyl-p-benzoquinone
- EDTA:
-
ethylenediamine tetraacetic acid
- EPR:
-
electron paramagnetic resonance
- Hepes:
-
N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid
- MES:
-
2-[N-Morpholino]ethanesulfonic acid
- OEE:
-
oxygen evolving enhancer
- PS II:
-
photosystem II
- SDS-PAGE:
-
sodium dedocyl sulfate polyacrylamide gel electrophoresis
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Shim, H., Cao, J., Govindjee et al. Purification of highly active oxygen-evolving photosystem II from Chlamydomonas reinhardtii . Photosynth Res 26, 223–228 (1990). https://doi.org/10.1007/BF00033135
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DOI: https://doi.org/10.1007/BF00033135