2021
In vitro fluorescence assay to measure GDP/GTP exchange of guanine nucleotide exchange factors of Rho family GTPases
Blaise AM, Corcoran EE, Wattenberg ES, Zhang YL, Cottrell JR, Koleske AJ. In vitro fluorescence assay to measure GDP/GTP exchange of guanine nucleotide exchange factors of Rho family GTPases. Biology Methods And Protocols 2021, 7: bpab024. PMID: 35087952, PMCID: PMC8789339, DOI: 10.1093/biomethods/bpab024.Peer-Reviewed Original ResearchNucleotide exchange factorsGEF domainExchange factorGuanine nucleotide exchange factorsGDP/GTP exchangeRho family GEFsActivation of GTPasesFluorescence-based analysisRare variantsGTP loadingGTP exchangeRho familyHigh-throughput screeningWhole-exome sequencingCellular activitiesGenetic variantsGTPasesGEFExome sequencingCritical roleUser-friendly protocolComplete understandingHuman developmentVariantsGEFs
2020
Scaffold association factor B (SAFB) is required for expression of prenyltransferases and RAS membrane association
Zhou M, Kuruvilla L, Shi X, Viviano S, Ahearn IM, Amendola CR, Su W, Badri S, Mahaffey J, Fehrenbacher N, Skok J, Schlessinger J, Turk BE, Calderwood DA, Philips MR. Scaffold association factor B (SAFB) is required for expression of prenyltransferases and RAS membrane association. Proceedings Of The National Academy Of Sciences Of The United States Of America 2020, 117: 31914-31922. PMID: 33257571, PMCID: PMC7749360, DOI: 10.1073/pnas.2005712117.Peer-Reviewed Original ResearchMeSH KeywordsAlkyl and Aryl TransferasesCell MembraneComputational BiologyCRISPR-Cas SystemsDatasets as TopicDimethylallyltranstransferaseGene Knockdown TechniquesHumansMatrix Attachment Region Binding ProteinsNeoplasmsNuclear Matrix-Associated ProteinsProtein PrenylationProtein SubunitsProto-Oncogene Proteins p21(ras)Receptors, EstrogenConceptsMembrane associationRAS membrane associationFarnesyltransferase inhibitorsPrenylation pathwayGenome-wide CRISPRGTP loadingAlternative prenylationMutant cellsNuclear proteinsKRAS membrane associationsRAS isoformsΑ-subunitGrowth inhibitionExpressionFactor BPathwayAnticancer therapyAlternative therapeutic strategiesPrenyltransferasesRasTherapeutic strategiesCRISPRFarnesyltransferaseMislocalizationPrenylation
2014
The Intraflagellar Transport Protein IFT27 Promotes BBSome Exit from Cilia through the GTPase ARL6/BBS3
Liew GM, Ye F, Nager AR, Murphy JP, Lee JS, Aguiar M, Breslow DK, Gygi SP, Nachury MV. The Intraflagellar Transport Protein IFT27 Promotes BBSome Exit from Cilia through the GTPase ARL6/BBS3. Developmental Cell 2014, 31: 265-278. PMID: 25443296, PMCID: PMC4255629, DOI: 10.1016/j.devcel.2014.09.004.Peer-Reviewed Original ResearchConceptsCargo entryBardet-Biedl syndrome proteinsIntraflagellar transport (IFT) machineryBiochemical reconstitution assaysNucleotide-free formCiliary exitTransport machinerySyndrome proteinGTP loadingUnbiased proteomicsCoat assemblyReconstitution assaysBBSomeARL6IFT27CiliaBBS3AssemblyProteomicsMachineryProteinCargoSortingExitActivationRegulation of Rac1 translocation and activation by membrane domains and their boundaries
Moissoglu K, Kiessling V, Wan C, Hoffman BD, Norambuena A, Tamm LK, Schwartz MA. Regulation of Rac1 translocation and activation by membrane domains and their boundaries. Journal Of Cell Science 2014, 127: 2565-2576. PMID: 24695858, PMCID: PMC4038948, DOI: 10.1242/jcs.149088.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsCell MembraneFluorescence Resonance Energy TransferGTPase-Activating ProteinsHEK293 CellsHumansMembrane MicrodomainsMiceNIH 3T3 CellsProtein BindingProtein Transportrac1 GTP-Binding Proteinrho-Specific Guanine Nucleotide Dissociation InhibitorsSignal TransductionUnilamellar LiposomesConceptsFluorescence resonance energy transferMembrane domainsRac1 translocationGDP dissociation inhibitor proteinLiquid-ordered membrane domainsGTPase-activating proteinsNon-raft regionsNon-raft domainsBinding of Rac1Activation of Rac1Single-molecule analysisGTP loadingRho GTPasesLipid raftsRac1 localizationRho GTPaseInhibitor proteinResult of inactivationRac1Resonance energy transferFunctional studiesNovel mechanismLipid bilayersTranslocationRafts
2000
Determination of GTP loading on Rho
Ren X, Schwartz M. Determination of GTP loading on Rho. Methods In Enzymology 2000, 325: 264-272. PMID: 11036609, DOI: 10.1016/s0076-6879(00)25448-7.Peer-Reviewed Original ResearchConceptsRho-binding domainGTP-RhoLow molecular weight GTPaseAffinity precipitation assaysActin cytoskeleton organizationGTP loadingCytoskeleton organizationWeight GTPaseGTPase activityRho effectorCell lysatesGTPaseRhoPrecipitation assaysTRBDWestern immunoblottingDomainQuality controlPositive controlAssaysRhotekinEffectorsProtein
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