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Electron transfer protein complexes in the thylakoid membranes of heterocysts from the cyanobacterium Nostoc punctiforme
Authors:Tanai Cardona  Pengpeng Zhang  Eva-Mari Aro  Ann Magnuson
Institution:a Department of Photochemistry and Molecular Science, Ångström laboratory, Uppsala University, P.O. Box 523, SE-75120 Uppsala, Sweden
b Plant Physiology and Molecular Biology, Department of Biology, University of Turku, FIN-20014 Turku, Finland
Abstract:Filamentous, heterocystous cyanobacteria are capable of nitrogen fixation and photoautotrophic growth. Nitrogen fixation takes place in heterocysts that differentiate as a result of nitrogen starvation. Heterocysts uphold a microoxic environment to avoid inactivation of nitrogenase, e.g. by downregulation of oxygenic photosynthesis. The ATP and reductant requirement for the nitrogenase reaction is considered to depend on Photosystem I, but little is known about the organization of energy converting membrane proteins in heterocysts. We have investigated the membrane proteome of heterocysts from nitrogen fixing filaments of Nostoc punctiforme sp. PCC 73102, by 2D gel electrophoresis and mass spectrometry. The membrane proteome was found to be dominated by the Photosystem I and ATP-synthase complexes. We could identify a significant amount of assembled Photosystem II complexes containing the D1, D2, CP43, CP47 and PsbO proteins from these complexes. We could also measure light-driven in vitro electron transfer from Photosystem II in heterocyst thylakoid membranes. We did not find any partially disassembled Photosystem II complexes lacking the CP43 protein. Several subunits of the NDH-1 complex were also identified. The relative amount of NDH-1M complexes was found to be higher than NDH-1L complexes, which might suggest a role for this complex in cyclic electron transfer in the heterocysts of Nostoc punctiforme.
Keywords:ATP  adenosine triphosphate  BN  blue native  Chl  chlorophyll  CLSM  confocal laser scanning microscopy  DM  d-maltoside" target="_blank">n-dodecyl β-d-maltoside  DBMIB  dibromothymoquinone  DCBQ  2  6-dichloro-p-benzoquinone  DCMU  3-(3  4-dichlorophenyl)-1  1-dimethylurea  DCPIP  2  6-dichlorophenolindophenol  DPC  diphenylcarbazide  EDTA  ethylene diamine tetraacetic acid  ESI  electrospray ionization  FNR  ferredoxin:NADP+ reductase  Hepes  4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid  Mes  4-morpholinoethane-sulfonic acid  MS  mass spectrometry  NDH-1  NAD(P)H dehydrogenase  PAGE  polyacrylamide gel electrophoresis  PPBQ  phenyl-p-benzoquinone  PSI  Photosystem I  PSII  Photosystem II  Rubisco  Ribulose-1  5-bisphosphate carboxylase/oxygenase  SDS  sodium dodecyl sulfate  Tris  tris(hydroxymethyl)aminomethane  TOF  time of flight
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