Separation of chlorophyll-protein complexes by Deriphat polyacrylamide gradient gel electrophoresis.
An improved Deriphat polyacrylamide gradient gel electrophoresis system was developed for the separation of chlorophyll-protein complexes. The relatively good resolution of the starting discontinuous gel system was further improved by using glycerol in gels and an acrylamide gradient with high acrylamide-to-N,N'-methylenebisacrylamide ratio in the separating gel. By applying mild but efficient glycosidic detergents for solubilization, and Deriphat to gels and buffers, the stability of complexes was increased, and only a low amount of pigment was removed. The advantage of our system is the better resolution of larger-size complexes, especially those of photosystem I. In addition, it makes possible an easier interpretation of results due to less overlapping of photosystem I and photosystem II bands when different plant species or the effects of different treatments are compared using whole thylakoid membranes.
Sárvári
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Department of Plant Physiology, Eötvös University, Budapest, Hungary.
Nyitrai
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D002734:Chlorophyll / Q000302:isolation & purification* / Q000378:metabolism; D004589:Electrophoresis, Disc / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D001467:Hordeum / Q000737:chemistry / Q000378:metabolism; D007202:Indicators and Reagents; D045342:Light-Harvesting Protein Complexes; D008970:Molecular Weight; D015388:Organelles / Q000737:chemistry / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000302:isolation & purification* / Q000378:metabolism; D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D014908:Triticum / Q000737:chemistry / Q000378:metabolism; D003313:Zea mays / Q000737:chemistry / Q000378:metabolism
D007202:Indicators and Reagents; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D002734:Chlorophyll