Abstract
LHC II isolated from carnation leaves has been solubilized and resolved by a newly developed, vertical-bed non-denaturing isoelectric focusing in polyacrylamide slab gels to yield three trimeric subcomplexes focusing at pH 4.52, 4.42 and 4.37 (designated a, b and c, respectively), comprising approximately 38%, 24% and 38% of the chlorophyll. The spectroscopic data demonstrated a close similarity among LHC II subcomplexes concerning their chlorophyll content and organization. The most alkaline and the most acidic subcomplex contained the 27 kDa polypeptide of LHC II while the intermediate pI fraction contained both LHC II polypeptides, i.e. 27 kDa and 26 kDa ones associated at 2:1 stoichiometry. The 27 kDa polypeptide could be resolved by denaturing isoelectrofocusing into 10 pI molecular isoforms covering 5.90–4.20 pH range. Three of the isoforms were found in the subcomplexes a and b and eight in the subcomplex c. The 26 kDa polypeptide comprised the unique pI molecular isoform focusing at pH 5.61.
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Abbreviations
- CBB G-250:
-
Coomassie Brilliant Blue G-250
- chl:
-
chlorophyll
- DM:
-
n-dodecyl-β-d-maltoside
- EDTA:
-
ethylendiaminotetraacetic acid
- IEF:
-
isoelectric focusing
- LHC II:
-
the main light-harvesting chlorophyll a/b-protein complex of Photosystem II
- LHCP II:
-
apoprotein of the main light-harvesting chlorophyll a/b-protein complex of Photosystem II
- NP-40:
-
polyethyleneglycol-p-isooctylphenyl ether
- pI:
-
isoelectric point
- OG:
-
octyl-β-d-glucopyranoside
- PS II:
-
Photosystem II
- SDS-PAGE:
-
sodium dodecylsulphate polyacrylamide gel electrophoresis
- TCA:
-
trichlorooacetic acid
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Jackowski, G., Przymusiński, R. The resolution and biochemical characterization of subcomplexes of the main light-harvesting chlorophyll a/b-protein complex of Photosystem II (LHC II). Photosynth Res 43, 41–48 (1995). https://doi.org/10.1007/BF00029461
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DOI: https://doi.org/10.1007/BF00029461