6533b852fe1ef96bd12aad0e
RESEARCH PRODUCT
Chimeric proteins tagged with specific 3xHA cassettes may present instability and functional problems
M. Carmen BañóEster MéndezInma QuilisSara Saiz-baggettoJ. Carlos Igualsubject
0301 basic medicinePhysiologyProtein Extractionlcsh:MedicineYeast and Fungal ModelsPolymerase Chain ReactionBiochemistryGreen fluorescent proteinEpitopesDatabase and Informatics MethodsGene Expression Regulation FungalImmune PhysiologyProtein purificationMacromolecular Structure AnalysisMedicine and Health SciencesProto-Oncogene Proteins c-myclcsh:ScienceStainingExtraction TechniquesImmune System ProteinsMultidisciplinarybiologyGene targetingProtein subcellular localization predictionMembrane StainingExperimental Organism SystemsGene TargetingArtifactsSequence AnalysisPlasmidsResearch ArticleProtein StructureSaccharomyces cerevisiae ProteinsBioinformaticsRecombinant Fusion ProteinsGenetic VectorsGreen Fluorescent ProteinsImmunologySaccharomyces cerevisiaeHemagglutinins ViralSaccharomyces cerevisiaeComputational biologyResearch and Analysis MethodsGreen Fluorescent ProteinGenomic InstabilityAntibodiesProtein–protein interactionProto-Oncogene Proteins c-mycSaccharomyces03 medical and health sciencesModel OrganismsAmino Acid Sequence AnalysisMolecular BiologyStaining and Labelinglcsh:ROrganismsFungiBiology and Life SciencesProteinsbiology.organism_classificationFusion proteinYeastLuminescent Proteins030104 developmental biologySpecimen Preparation and Treatmentlcsh:QProtein Structure Networksdescription
Epitope-tagging of proteins has become a widespread technique for the analysis of protein function, protein interactions and protein localization among others. Tagging of genes by chromosomal integration of PCR amplified cassettes is a widely used and fast method to label proteins in vivo. Different systems have been developed during years in the yeast Saccharomyces cerevisiae. In the present study, we analysed systematically a set of yeast proteins that were fused to different tags. Analysis of the tagged proteins revealed an unexpected general effect on protein level when some specific tagging module was used. This was due in all cases to a destabilization of the proteins and caused a reduced protein activity in the cell that was only apparent in particular conditions. Therefore, an extremely cautious approach is required when using this strategy.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2017-05-17 | PLOS ONE |