Combining laser capture microdissection and proteomics reveals an active translation machinery controlling invadosome formation
Nat Commun. 2018-05-23; 9(1):
DOI: 10.1038/s41467-018-04461-9
Read on PubMed
1. 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.
Ezzoukhry Z(1)(2)(3), Henriet E(1)(2), Cordelières FP(2)(4), Dupuy JW(2)(5),
Maître M(6), Gay N(1)(2), Di-Tommaso S(1)(2), Mercier L(7)(8)(9), Goetz
JG(7)(8)(9), Peter M(10), Bard F(11), Moreau V(1)(2), Raymond AA(1)(2)(12),
Saltel F(13)(14)(15).
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. .
(14)Université de Bordeaux, 146 Rue Léo Saignat, Bordeaux, F-33076, France.
.
(15)Oncoprot, INSERM UMR1053-TBM Core US005, 146 Rue Léo Saignat, Bordeaux,
F-33000, France. .
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.
DOI: 10.1038/s41467-018-04461-9
PMCID: PMC5966458
PMID: 29795195