0000000000997265

AUTHOR

Swati Tyagi

showing 3 related works from this author

Comparative analysis of the coordinated motion of Hsp70s from different organelles observed by single-molecule three-color FRET.

2021

Cellular function depends on the correct folding of proteins inside the cell. Heat-shock proteins 70 (Hsp70s), being among the first molecular chaperones binding to nascently translated proteins, aid in protein folding and transport. They undergo large, coordinated intra- and interdomain structural rearrangements mediated by allosteric interactions. Here, we applied a three-color single-molecule Forster resonance energy transfer (FRET) combined with three-color photon distribution analysis to compare the conformational cycle of the Hsp70 chaperones DnaK, Ssc1, and BiP. By capturing three distances simultaneously, we can identify coordinated structural changes during the functional cycle. Be…

chemistry.chemical_classificationOrganellesMultidisciplinarySaccharomyces cerevisiae ProteinsAllosteric regulationPeptideSaccharomyces cerevisiaeBiological SciencesMitochondrial Membrane Transport ProteinsRecombinant ProteinsSingle Molecule ImagingFolding (chemistry)Förster resonance energy transferchemistryHeat shock proteinBiophysicsEscherichia coliFluorescence Resonance Energy TransferMoleculeProtein foldingNucleotideHSP70 Heat-Shock ProteinsMolecular ChaperonesProceedings of the National Academy of Sciences of the United States of America
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Two differential binding mechanisms of FG-nucleoporins and nuclear transport receptors

2018

Summary Phenylalanine-glycine-rich nucleoporins (FG-Nups) are intrinsically disordered proteins, constituting the selective barrier of the nuclear pore complex (NPC). Previous studies showed that nuclear transport receptors (NTRs) were found to interact with FG-Nups by forming an “archetypal-fuzzy” complex through the rapid formation and breakage of interactions with many individual FG motifs. Here, we use single-molecule studies combined with atomistic simulations to show that, in sharp contrast, FG-Nup214 undergoes a coupled reconfiguration-binding mechanism when interacting with the export receptor CRM1. Association and dissociation rate constants are more than an order of magnitude lowe…

0301 basic medicineModels MolecularGlycosylationglycosylationProtein ConformationPhenylalanineGlycineSequence (biology)Intrinsically disordered proteinsnuclear transport receptorssingle-molecule FRETGeneral Biochemistry Genetics and Molecular BiologyArticle03 medical and health scienceschemistry.chemical_compound0302 clinical medicineEscherichia coliFluorescence Resonance Energy TransferHumansNuclear poreReceptorlcsh:QH301-705.5Single-molecule FRETmolecular dynamics simulationsbinding mechanismintrinsically disordered proteinFG-Nup3. Good healthNuclear Pore Complex Proteins030104 developmental biologychemistrylcsh:Biology (General)BiophysicsNuclear PoreNucleoporinNuclear transport030217 neurology & neurosurgeryProtein BindingCell Reports
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Precision and accuracy of single-molecule FRET measurements-a multi-laboratory benchmark study

2018

Single-molecule Forster resonance energy transfer (smFRET) is increasingly being used to determine distances, structures, and dynamics of biomolecules in vitro and in vivo. However, generalized protocols and FRET standards to ensure the reproducibility and accuracy of measurements of FRET efficiencies are currently lacking. Here we report the results of a comparative blind study in which 20 labs determined the FRET efficiencies (E) of several dye-labeled DNA duplexes. Using a unified, straightforward method, we obtained FRET efficiencies with s.d. between +/- 0.02 and +/- 0.05. We suggest experimental and computational procedures for converting FRET efficiencies into accurate distances, and…

0301 basic medicinePHOTON DISTRIBUTIONDYNAMICSAccuracy and precisionTechnologyBiophysicsRESONANCE ENERGY-TRANSFERBiochemistryMedical and Health SciencesArticle03 medical and health sciencesBlind studySingle-molecule biophysicsALTERNATING-LASER EXCITATIONSTRUCTURAL INFORMATIONFluorescence resonance energy transferDEPENDENCEQuantitative assessmentLife ScienceFLUORESCENCEStructure determinationMolecular BiologyQCVLAGBiophysical methodsReproducibilityReproducibility of ResultsCell BiologySingle-molecule FRETDNABiological SciencesPublisher CorrectionQPSPECTROSCOPIC RULER030104 developmental biologyFörster resonance energy transferBiofysicaBenchmark (computing)Photon distributionEPSREFRACTIVE-INDEXLaboratoriesBiological systemBiotechnologyDevelopmental Biology
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