Resolution of 16 to 20 chlorophyll-protein complexes using a low ionic strength native green gel system.
Conventional native "green gel" systems resolve at most 10 chlorophyll-protein complexes from thylakoid membranes of higher plants and green algae. Such analyses suggest a simplicity of the thylakoid membrane that is not supported by a growing body of evidence on the heterogeneity of photosystems I and II (PSI and PSII) and their associated antennae (LHCI and LHCII). We report here the development and characterization of a low ionic strength native "green gel" system that resolves from 16 to 20, mostly large chlorophyll-protein complexes from a variety of higher plant and green algal species with very little release of free pigment. In Chlamydomonas, this system resolves multiple PSI-LHCI complexes, multiple PSII-LHCII complexes, four oligomeric LHCII complexes, as well as several low electrophoretic mobility reaction center complexes, and a number of small complexes. We have obtained similar resolution with a large number of higher plant and green algal species. We also demonstrate how this system can be used as a sort of "fingerprinting" technique to distinguish thylakoids of different species, and for the analysis of photosynthetic mutants, using the chlorophyll b-less chlorina f2 mutant of barley as an example.
Allen
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KD
Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder 80309.
Staehelin
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LA
D002734:Chlorophyll / Q000032:analysis / Q000302:isolation & purification*; D003902:Detergents; D004586:Electrophoresis / Q000379:methods; D005782:Gels; D007700:Kinetics; D046911:Macromolecular Substances; D008566:Membranes / Q000737:chemistry; D009154:Mutation; D009994:Osmolar Concentration; D010940:Plant Proteins / Q000302:isolation & purification*
D003902:Detergents; D005782:Gels; D046911:Macromolecular Substances; D010940:Plant Proteins; D002734:Chlorophyll; C037184:chlorophyll b