Label-free quantitative proteomic analysis of the YAP/TAZ interactome
| Autor: | Bernhard Schermer, Sandra Habbig, Thomas Benzing, Tobias Lamkemeyer, Caroline Pahmeyer, Markus M. Rinschen, Priyanka Kohli, Malte P. Bartram |
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| Rok vydání: | 2014 |
| Předmět: |
Proteomics
Physiology Recombinant Fusion Proteins Protein subunit Green Fluorescent Proteins Molecular Sequence Data Cell Cycle Proteins Computational biology Protein Serine-Threonine Kinases Biology Transfection Interactome law.invention Mice Tandem Mass Spectrometry Transcription (biology) law Protein Interaction Mapping Animals Cluster Analysis Humans Immunoprecipitation Hippo Signaling Pathway Amino Acid Sequence Protein Interaction Maps Transcription factor Adaptor Proteins Signal Transducing Label free Genetics Effector Computational Biology YAP-Signaling Proteins Cell Biology Phosphoproteins HEK293 Cells Gene Expression Regulation NIH 3T3 Cells Suppressor Acyltransferases Protein Binding Signal Transduction Transcription Factors |
| Zdroj: | American Journal of Physiology-Cell Physiology. 306:C805-C818 |
| ISSN: | 1522-1563 0363-6143 |
| DOI: | 10.1152/ajpcell.00339.2013 |
| Popis: | The function of an individual protein is typically defined by protein-protein interactions orchestrating the formation of large complexes critical for a wide variety of biological processes. Over the last decade the analysis of purified protein complexes by mass spectrometry became a key technique to identify protein-protein interactions. We present a fast and straightforward approach for analyses of interacting proteins combining a Flp-in single-copy cellular integration system and single-step affinity purification with single-shot mass spectrometry analysis. We applied this protocol to the analysis of the YAP and TAZ interactome. YAP and TAZ are the downstream effectors of the mammalian Hippo tumor suppressor pathway. Our study provides comprehensive interactomes for both YAP and TAZ and does not only confirm the majority of previously described interactors but, strikingly, revealed uncharacterized interaction partners that affect YAP/TAZ TEAD-dependent transcription. Among these newly identified candidates are Rassf8, thymopoetin, and the transcription factors CCAAT/enhancer-binding protein (C/EBP)β/δ and core-binding factor subunit β (Cbfb). In addition, our data allowed insights into complex stoichiometry and uncovered discrepancies between the YAP and TAZ interactomes. Taken together, the stringent approach presented here could help to significantly sharpen the understanding of protein-protein networks. |
| Databáze: | OpenAIRE |
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