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Molecular Basis of the Mechanisms Controlling MASTL.

Molecular & cellular proteomics : MCP (2019-12-20)
Dario Hermida, Gulnahar B Mortuza, Anna-Kathrine Pedersen, Irina Pozdnyakova, Tam T T N Nguyen, Maria Maroto, Michael Williamson, Tasja Ebersole, Giuseppe Cazzamali, Kasper Rand, Jesper V Olsen, Marcos Malumbres, Guillermo Montoya
RÉSUMÉ

The human MASTL (Microtubule-associated serine/threonine kinase-like) gene encodes an essential protein in the cell cycle. MASTL is a key factor preventing early dephosphorylation of M-phase targets of Cdk1/CycB. Little is known about the mechanism of MASTL activation and regulation. MASTL contains a non-conserved insertion of 550 residues within its activation loop, splitting the kinase domain, and making it unique. Here, we show that this non-conserved middle region (NCMR) of the protein is crucial for target specificity and activity. We performed a phosphoproteomic assay with different MASTL constructs identifying key phosphorylation sites for its activation and determining whether they arise from autophosphorylation or exogenous kinases, thus generating an activation model. Hydrogen/deuterium exchange data complements this analysis revealing that the C-lobe in full-length MASTL forms a stable structure, whereas the N-lobe is dynamic and the NCMR and C-tail contain few localized regions with higher-order structure. Our results indicate that truncated versions of MASTL conserving a cryptic C-Lobe in the NCMR, display catalytic activity and different targets, thus establishing a possible link with truncated mutations observed in cancer-related databases.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
MBP déphosphorylée
Sigma-Aldrich
Anti-Mastl Antibody, clone 4F9, clone 4F9, from mouse