Structure-based optimization of a PDZ binding motif within a viral peptide stimulates neurite outgrowth

Zakir Khan (1) , Elouan Terrien (2) , Florent Delhommel (2) , Cynthia Lefebvre-Omar (3) , Delphine Bohl (3) , Sandrine Vitry (1) , Clara Bernard (1) , Juan Morua Ramirez (2) , Alain Chaffotte (2) , Kevin Ricquier (2) , Renaud Vincentelli (4) , Henri Buc (5) , Christophe Préhaud (1) , Nicolas Wolff (2) , Monique Lafon (1)

1 Neuro-Immunologie Virale - Viral Neuro-immunology
2 Résonance Magnétique Nucléaire des Biomolécules
3 ICM - Institut du Cerveau = Paris Brain Institute
4 AFMB - Architecture et fonction des macromolécules biologiques
5 IP - Institut Pasteur [Paris]

Protection of neuronal homeostasis is a major goal in the management of neurodegenerative diseases. Microtubule-associated Ser/Thre kinase 2 (MAST2), inhibits neurite outgrowth, and its inhibition therefore represents a potential therapeutic strategy. We previously reported that a viral protein (G-protein from rabies virus) capable of interfering with protein-protein interactions between the PDZ domain of MAST2 and the C-terminal moieties of its cellular partners counteracts MAST2-mediated suppression of neurite outgrowth. Here, we designed peptides derived from the native viral protein to increase the affinity of these peptides for the MAST2 PDZ domain. Our strategy involved modifying the length and flexibility of the non-interacting sequence linking the two subsites anchoring the peptide to the PDZ domain. Three peptides, Neurovita1 (NV1), NV2, and NV3, were selected, and we found that they all had increased affinities for the MAST2 PDZ domain, with Kd values decreasing from 1300 to 60 nM, while target selectivity was maintained. A parallel biological assay evaluating neurite extension and branching in cell cultures revealed that the NV peptides gradually improved neural activity, with the efficacies of these peptides for stimulating neurite outgrowth mirroring their affinities for MAST2-PDZ. We also show that NVs can be delivered into the cytoplasm of neurons as a gene or peptide. In summary, our findings indicate that virus-derived peptides targeted to MAST2-PDZ stimulate neurite outgrowth in several neuron types, opening up promising avenues for potentially using NVs in the management of neurodegenerative diseases.

Mots clés: neurite outgrowth; protein-protein interaction; neurodegenerative diseases; serine/threonine protein kinase; structure function analysis; therapeutical drug candidate; Nuclear Magnetic Resonance; viral protein; Interfering peptide; PDZ domain

Domaines: Sciences cognitives; Neurosciences

Zakir Khan, Elouan Terrien, Florent Delhommel, Cynthia Lefebvre-Omar, Delphine Bohl, et al.. Structure-based optimization of a PDZ binding motif within a viral peptide stimulates neurite outgrowth. Journal of Biological Chemistry, 2019, 294 (37), pp.137555-13768. ⟨10.1074/jbc.RA119.008238⟩. ⟨pasteur-02271324⟩