2018
Multiplexed Real-Time NMR GTPase Assay for Simultaneous Monitoring of Multiple Guanine Nucleotide Exchange Factor Activities from Human Cancer Cells and Organoids
Gebregiworgis T, Marshall C, Nishikawa T, Radulovich N, Sandí M, Fang Z, Rottapel R, Tsao M, Ikura M. Multiplexed Real-Time NMR GTPase Assay for Simultaneous Monitoring of Multiple Guanine Nucleotide Exchange Factor Activities from Human Cancer Cells and Organoids. Journal Of The American Chemical Society 2018, 140: 4473-4476. PMID: 29543440, DOI: 10.1021/jacs.7b13703.Peer-Reviewed Original ResearchConceptsGuanine nucleotide exchange factorsRhoA-specific Guanine Nucleotide Exchange FactorGuanine nucleotide exchange factor activityNucleotide exchange factor activityHigher GEF activityNucleotide exchange factorsImportant cellular functionsExchange factor activityExchange of GTPProtein of interestReal-time NMRRNA-seq dataCultured human cellsGEF assaysHuman cancer cellsSmall GTPasesExchange factorGEF activityCellular functionsIsotopic labeling schemesNucleotide exchangeGEF-H1GTPase assaysFunctional analysisHuman cells
2016
Biochemical Classification of Disease-associated Mutants of RAS-like Protein Expressed in Many Tissues (RIT1)*
Fang Z, Marshall C, Yin J, Mazhab-Jafari M, Gasmi-Seabrook G, Smith M, Nishikawa T, Xu Y, Neel B, Ikura M. Biochemical Classification of Disease-associated Mutants of RAS-like Protein Expressed in Many Tissues (RIT1)*. Journal Of Biological Chemistry 2016, 291: 15641-15652. PMID: 27226556, PMCID: PMC4957048, DOI: 10.1074/jbc.m116.714196.Peer-Reviewed Original ResearchConceptsDisease-associated mutationsGTPase cycleIntrinsic GTP hydrolysis rateA57GRIT1 mutationsIntrinsic nucleotide exchange rateDisease-associated mutantsGTP hydrolysis rateNucleotide exchange rateReal-time NMRSmall guanosine triphosphataseHEK293T cellsPulldown assaysGuanosine triphosphataseGTP hydrolysisNoonan syndromeGTPase assaysDivergent residuesRIT1GTPase propertiesMutantsModest activationH-RASMutationsUrinary tract cancer