Deciphering the unconventional peptide binding to the PDZ domain of MAST2.
Phosphatase and tensin homologue (PTEN) and microtubule-associated serine threonine kinase 2 (MAST2) are key negative regulators of survival pathways in neuronal cells. The two proteins interact via the PDZ (PSD-95, Dlg1, Zo-1) domain of MAST2 (MAST2-PDZ). During infection by rabies virus, the viral glycoprotein competes with PTEN for interaction with MAST2-PDZ and promotes neuronal survival. The C-terminal PDZ-binding motifs (PBMs) of the two proteins bind similarly to MAST2-PDZ through an unconventional network of connectivity involving two anchor points. Combining stopped-flow fluorescence, analytical ultracentrifugation (AUC), microcalorimetry and NMR, we document the kinetics of interaction between endogenous and viral ligands to MAST2-PDZ as well as the dynamic and structural effects of these interactions. Viral and PTEN peptide interactions to MAST2-PDZ occur via a unique kinetic step which involves both canonical C-terminal PBM binding and N-terminal anchoring. Indirect effects induced by the PBM binding include modifications to the structure and dynamics of the PDZ dimerization surface which prevent MAST2-PDZ auto-association. Such an energetic communication between binding sites and distal surfaces in PDZ domains provides interesting clues for protein regulation overall.
Département de Biologie Structurale et Chimie, Unité de Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur, F-75724 Paris, France UMR 3528, CNRS, F-75724 Paris, France Université Pierre et Marie Curie, Cellule Pasteur UPMC, rue du Docteur Roux, 75015 Paris, France.
Florent
F
Delhommel
Département de Biologie Structurale et Chimie, Unité de Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur, F-75724 Paris, France UMR 3528, CNRS, F-75724 Paris, France.
Alain
A
Chaffotte
Département de Biologie Structurale et Chimie, Unité de Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur, F-75724 Paris, France UMR 3528, CNRS, F-75724 Paris, France.
Elouan
E
Terrien
Plate-Forme de Biophysique des Macromolécules, Institut Pasteur, F-75724 Paris, France UMR 3528, CNRS, F-75724 Paris, France.
Bertrand
B
Raynal
Invited Scientist, Institut Pasteur F-75724 Paris, France.
Henri
H
Buc
Département de Biologie Structurale et Chimie, Unité de Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur, F-75724 Paris, France UMR 3528, CNRS, F-75724 Paris, France.
Muriel
M
Delepierre
Département de Biologie Structurale et Chimie, Unité de Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur, F-75724 Paris, France UMR 3528, CNRS, F-75724 Paris, France.
Florence
F
Cordier
Département de Biologie Structurale et Chimie, Unité de Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur, F-75724 Paris, France UMR 3528, CNRS, F-75724 Paris, France nicolas.wolff@pasteur.fr.
Nicolas
N
Wolff
D006801:Humans; D008869:Microtubule-Associated Proteins / Q000737:chemistry* / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D055503:Protein Multimerization; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000378:metabolism; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D011820:Rabies virus / Q000737:chemistry* / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
D008869:Microtubule-Associated Proteins; D014764:Viral Proteins; C503880:MAST2 protein, human; D017346:Protein Serine-Threonine Kinases
PDZ domains; intradomain energetic connectivity; pre-steady-state kinetics; protein–protein interactions