Combining laser capture microdissection and proteomics reveals an active translation machinery controlling invadosome formation

• DOI: 10.1038/s41467-018-04461-9
• Publication Date: 2018-05-17T10:11:25Z
• Authors: Zakaria Ezzoukhry; Elodie Henriet; Fabrice P. Cordel...; Jean-William Dupuy; Marlène Maître; Nathan Gay; Sylvaine Di-Tommaso; Luc Mercier; Jacky G. Goetz; Marion Peter; Frédéric Bard; Violaine Moreau; Anne-Aurélie Raymond; Frédéric Saltel
• Keywords: Laser capture microdissection; Proteome; Microdissection

Invadosomes are F-actin-based structures involved in extracellular matrix degradation, cell invasion, and metastasis formation. Analyzing their proteome is crucial to decipher molecular composition, understand mechanisms, find specific elements target them. However, the analysis of invadosomes challenging, because it difficult maintain integrity during isolation. In addition, classical purification methods often suffer from contaminations, which may impair data validation. To ensure identification invadosome components, we here develop a method that combines laser microdissection mass spectrometry, enabling subcellular native state based on low amounts input material. Using this combinatorial method, show contain components translational machinery, addition known marker proteins. Moreover, functional validation reveals protein translation activity an inherent property invadosomes, required structure activity.