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Nat Commun. 2018 May 23;9(1):2031. doi: 10.1038/s41467-018-04461-9.

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

Author information

1
INSERM, UMR1053, BaRITOn Bordeaux Research in Translational Oncology, 146 Rue Léo Saignat, Bordeaux, F-33000, France.
2
Université de Bordeaux, 146 Rue Léo Saignat, Bordeaux, F-33076, France.
3
Mohammed VI University of Health Sciences (UM6SS), Casablanca, Morocco.
4
Bordeaux Imaging Center, UMS 3420 CNRS-Université de Bordeaux-US4 INSERM, Pôle d'imagerie photonique, 146 Rue Léo Saignat, Bordeaux, F-33000, France.
5
Plateforme Protéome, Centre de Génomique Fonctionnelle, 146 Rue Léo Saignat, Bordeaux, F-33000, France.
6
Neurocentre Magendie U1215, INSERM, 146 Rue Léo Saignat, Bordeaux, F-33000, France.
7
Inserm U1109 MN3T, 1 Place de L'Hôpital, Strasbourg, F-67000, France.
8
LabEx Medalis, Université de Strasbourg, Strasbourg, F-67000, France.
9
Fédération de Médecine Translationnelle de Strasbourg (FMTS), Strasbourg, F-67000, France.
10
IGMM, CNRS, Univ. Montpellier, 1919 Route de Mende, Montpellier, F-34090, France.
11
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore.
12
Oncoprot, INSERM UMR1053-TBM Core US005, 146 Rue Léo Saignat, Bordeaux, F-33000, France.
13
INSERM, UMR1053, BaRITOn Bordeaux Research in Translational Oncology, 146 Rue Léo Saignat, Bordeaux, F-33000, France. frederic.saltel@inserm.fr.
14
Université de Bordeaux, 146 Rue Léo Saignat, Bordeaux, F-33076, France. frederic.saltel@inserm.fr.
15
Oncoprot, INSERM UMR1053-TBM Core US005, 146 Rue Léo Saignat, Bordeaux, F-33000, France. frederic.saltel@inserm.fr.

Abstract

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.

PMID:
29795195
PMCID:
PMC5966458
DOI:
10.1038/s41467-018-04461-9
[Indexed for MEDLINE]
Free PMC Article

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