深圳中恒华发股份有限公司.pptxVIP

Protein InteractionsMichel DumontierBlueprint Initiativemjdumont@ Lecture 4.1OutlineMolecular interactionsDiscoveryExperimentalComputationalStorageData MiningFuture DirectionsLecture 4.1BAMolecular InteractionsBetween two molecular objectsDNA, RNA, gene, protein, molecular complex, small molecule, photonBinding SitesUnder an Experimental ConditionWith a particular Cellular LocationPossibly causing some Chemical ActionLecture 4.1MassSpectrometryMicroarrayTwo-HybridGeneticsInteraction DiscoveryExpression, Interaction Data, Function, Protein modificationsLecture 4.1A measure of confidence?How do you know if the interaction really exists? Each method has its advantages and disadvantages. Be aware of systematic errors (i.e. tag effects)Be aware of contaminating proteins.Each method observes interactions from a slightly different experimental condition.Support from many different sources is certainly better than just one.Lecture 4.1High-throughput Mass Spectrometric Protein Complex Identification (HMS-PCI)Mike Tyers, SLRISte12Ho et al. Nature. 2002 Jan 10;415(6868):180-3Lecture 4.1FilteringRemove promiscuously binding proteins:Proteins identified by MS in control lanesProteins that were found at a high frequency in the overall experimentKnown high-abundant proteins ribosomal elementsLecture 4.1Synthetic Genetic InteractionsSynthetic genetic interactions (lethal, slow growth)Mate two mutants without phenotypes to get a daughter cell with a phenotypeSynthetic lethal (SL), slow growthrobotic mating using the yeast deletion libraryGenetic interactions provide functional data on protein interactions or redundant genesAbout 23% of known SLs (1295 - YPD+MIPS) are known protein interactions in yeastTong et al. Science. 2001 Dec 14;294(5550):2364-8Lecture 4.1Working overtimeCharlie Boone’s RobotsLecture 4.1Cell PolarityCell Wall Maintenance Cell StructureMitosisChromosome StructureDNA Synthesis DNA RepairUnknownOthersSynthetic Genetic Interactions in YeastTong, BoonePreBIND – Li