Overexpression and reconstitution of a Rieske iron-sulfur protein from the cyanobacterium Synechocystis PCC 6803.

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dc.contributor.authorSchneider, Den_US
dc.contributor.authorJaschkowitz, Ken_US
dc.contributor.authorSeidler, Aen_US
dc.contributor.authorRögner, Men_US
dc.date.accessioned2000-12-18en_US
dc.date.accessioned2009-05-27T09:35:51Z
dc.date.available2000-12-18en_US
dc.date.available2009-05-27T09:35:51Z
dc.date.issued2000-12-18en_US
dc.description.abstractChloroplast cyt b6f complexes as well as mitochondrial and bacterial cyt bc1 complexes contain a high potential Rieske iron-sulfur protein which is essential for their function. To characterise the isolated Rieske protein from the mesophilic cyanobacterium Synechocystis PCC6803 we cloned the encoding gene into an expression vector and overexpressed the protein in E. coli. In cells overexpressing the protein no typical Rieske type EPR signal was detected neither in membranes nor in inclusion bodies where the majority of the protein was deposited. The inclusion bodies were isolated from the E. coli cells and denaturated with 8 M urea. With a single anion exchange chromatographic step a pure protein could be obtained which was used for further experiments. The NifS like protein IscS was recently reported to mediate the incorporation of iron-sulfur clusters into ferredoxin in vitro. We used the recombinant IscS protein for the incorporation of the cluster into the folded Rieske apoprotein. Spectroscopic characterisation of the resultant protein by CD and EPR spectroscopy showed the presence of a typical Rieske iron-sulfur centre.en_US
dc.description.affiliationLehrstuhl für Biochemie der Pflanzen, Fakultät für Biologie, Ruhr-Universität Bochum, D-447800 Bochum, Germany.en_US
dc.identifier.citationSchneider D, Jaschkowitz K, Seidler A, Rögner M. Overexpression and reconstitution of a Rieske iron-sulfur protein from the cyanobacterium Synechocystis PCC 6803. Indian Journal of Biochemistry & Biophysics. 2000 Dec; 37(6): 441-6en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/27985
dc.language.isoengen_US
dc.source.urihttps://https://www.niscair.res.in/ScienceCommunication/ResearchJournals/rejour/ijbb/ijbb0.aspen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCloning, Molecularen_US
dc.subject.meshCyanobacteria --chemistryen_US
dc.subject.meshDNA Primersen_US
dc.subject.meshElectron Spin Resonance Spectroscopyen_US
dc.subject.meshElectron Transport Complex IIIen_US
dc.subject.meshIron-Sulfur Proteins --chemistryen_US
dc.subject.meshRecombinant Proteins --chemistryen_US
dc.titleOverexpression and reconstitution of a Rieske iron-sulfur protein from the cyanobacterium Synechocystis PCC 6803.en_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
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