Tryptophan fluorescence quenching as a binding assay to monitor protein conformation changes in the membrane of intact mitochondria

cg.contactH.SHARMA@CGIAR.ORGen_US
cg.contributor.centerInternational Crops Research Institute for the Semi-Arid Tropics - ICRISATen_US
cg.contributor.centerGulbarga Universityen_US
cg.contributor.crpCGIAR Research Program on Grain Legumes - GLen_US
cg.contributor.funderNot Applicableen_US
cg.coverage.countryINen_US
cg.coverage.regionSouthern Asiaen_US
cg.date.embargo-end-date2017-06-30en_US
cg.identifier.doihttps://dx.doi.org/10.1007/s10863-016-9653-0en_US
cg.isijournalISI Journalen_US
cg.issn1573-6881en_US
cg.issue3en_US
cg.journalJournal of Bioenergetics and Biomembranesen_US
cg.subject.agrovocagricultureen_US
cg.subject.agrovocmitochondriaen_US
cg.subject.agrovoccytochrome cen_US
cg.volume48en_US
dc.contributorSreeramulu, Kurubaen_US
dc.contributorSharma, Hari C.en_US
dc.creatorAkbar, Mohammad S.en_US
dc.date.accessioned2017-04-08T22:15:23Z
dc.date.available2017-04-08T22:15:23Z
dc.description.abstractIntrinsic protein fluorescence is due to aromatic amino acids, mainly tryptophan, which can be selectively measured by exciting at 295 nm. Changes in emission spectra of tryptophan are due to the protein conformational transitions, subunit association, ligand binding or denaturation, which affect the local environment surrounding the indole ring. In this study, tryptophan fluorescence was monitored in intact mitochondria at 333 nm following excitation at 295 nm in presence of insecticides using spectrofluorometer. Methylparathion, carbofuran, and endosulfan induced Trp fluorescence quenching and release of cytochrome c when incubated with the mitochondria, except fenvalarate. Mechanism of insecticide-induced mitochondrial toxicity for the tested insecticides has been discussed. Reduction in the intensity of tryptophan emission spectra of mitochondrial membrane proteins in presence of an increasing concentration of a ligand can be used to study the interaction of insecticides/drugs with the intact mitochondria. Furthermore, this assay can be readily adapted for studying protein–ligand interactions in intact mitochondria and in other cell organelles extending its implications for pesticide and pharma industry and in drug discovery.en_US
dc.formatPDFen_US
dc.identifierhttp://oar.icrisat.org/id/eprint/9365en_US
dc.identifierhttps://mel.cgiar.org/reporting/downloadmelspace/hash/fHZTAaTB/v/1277e2ee2eb6db9fc4c11b337632c620en_US
dc.identifier.citationMohammad S. Akbar, Kuruba Sreeramulu, Hari C. Sharma. (30/6/2016). Tryptophan fluorescence quenching as a binding assay to monitor protein conformation changes in the membrane of intact mitochondria. Journal of Bioenergetics and Biomembranes, 48 (3), pp. 241-7.en_US
dc.identifier.statusLimited accessen_US
dc.identifier.urihttps://hdl.handle.net/20.500.11766/6660
dc.languageenen_US
dc.publisherSpringer Verlag (Germany)en_US
dc.rightsCC-BY-NC-4.0en_US
dc.sourceJournal of Bioenergetics and Biomembranes;48,(2016) Pagination 241,7en_US
dc.subjecttrp fluorescence quenchingen_US
dc.subjectinsecticideen_US
dc.titleTryptophan fluorescence quenching as a binding assay to monitor protein conformation changes in the membrane of intact mitochondriaen_US
dc.typeJournal Articleen_US
dcterms.available2016-06-30en_US
dcterms.extent241-7en_US
mel.impact-factor2.080en_US

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