Combining laser capture microdissection and proteomics reveals an active translation machinery controlling invadosome formation.
Fiche publication
Date publication
mai 2018
Journal
Nature communications
Auteurs
Membres identifiés du Cancéropôle Est :
Dr GOETZ Jacky
Tous les auteurs :
Ezzoukhry Z, Henriet E, Cordelières FP, Dupuy JW, Maître M, Gay N, Di-Tommaso S, Mercier L, Goetz JG, Peter M, Bard F, Moreau V, Raymond AA, Saltel F
Lien Pubmed
Résumé
Invadosomes are F-actin-based structures involved in extracellular matrix degradation, cell invasion, and metastasis formation. Analyzing their proteome is crucial to decipher their molecular composition, to understand their mechanisms, and to find specific elements to target them. However, the specific analysis of invadosomes is challenging, because it is difficult to maintain their integrity during isolation. In addition, classical purification methods often suffer from contaminations, which may impair data validation. To ensure the specific identification of invadosome components, we here develop a method that combines laser microdissection and mass spectrometry, enabling the analysis of subcellular structures in their native state based on low amounts of input material. Using this combinatorial method, we show that invadosomes contain specific components of the translational machinery, in addition to known marker proteins. Moreover, functional validation reveals that protein translation activity is an inherent property of invadosomes, which is required to maintain invadosome structure and activity.
Mots clés
Actins, metabolism, Animals, Biomarkers, Tumor, analysis, Cell Line, Tumor, Chromatography, High Pressure Liquid, methods, Extracellular Matrix, metabolism, Humans, Laser Capture Microdissection, methods, Mice, NIH 3T3 Cells, Neoplasms, diagnosis, Podosomes, metabolism, Protein Biosynthesis, Proteomics, methods, RNA, Messenger, metabolism, Tandem Mass Spectrometry, methods
Référence
Nat Commun. 2018 May 23;9(1):2031