Computational and Experimental Identification of putative αTAT1 Modulators: Implications for Nervous System Function

Oksana Rybachuk^1,2Alexey Rayevsky^3,4*Mariia Stykhylias⁵Dariia Samofalova^5,6Elihja Bulgakov^5,7Maxim Platonov^4,7Yaroslav B. Blume⁵Pavel Karpov⁵

• ¹Institut Fiziologii imeni O O Bogomolca Nacional'na akademia nauk Ukraini, Kyiv, Ukraine
• ²DU Nacional'nij naukovij centr Institut kardiologii klinicnoi ta regenerativnoi medicini imeni akademika M D Strazeska NAMN Ukraini, Kyiv, Ukraine
• ³Institute of Food Biotechnology and Genomics, National Academy of Sciences of Ukraine (NAN Ukraine), Kyiv, Ukraine
• ⁴Institut molekularnoi biologii i genetiki Nacional'na akademia nauk Ukraini, Kyiv, Ukraine
• ⁵Institut harcovoi biotehnologii ta genomiki Nacional'na akademia nauk Ukraini, Kyiv, Ukraine
• ⁶Life Chemicals Kyiv, Kyiv, Ukraine
• ⁷Enamine Ltd, Kyiv, Ukraine

The project's primary objective is to understand how enzymes responsible for post-translational modifications (PTMs) of microtubule elements influence ion channels in excitatory peripheral nervous system (PNS) cells, and to subsequently identify potential pharmacological agents that can act upon these molecular targets. Having identified HDAC6 and αTAT1 as the most relevant proteins for further study, we designed a corresponding pharmacophore model hypothesis of ligand-dependent inhibition for αTAT1, which plays a key role in regulating microtubule dynamics within the cell. αTAT1 is the sole mammalian enzyme known to acetylate microtubules, a process associated with regulating the dynamics and protecting long-lived microtubules from mechanical stress. Additionally, αTAT1 plays a role in nuclear processes such as DNA replication, cell migration, and axonal transport. Given the importance of αTAT1 in cytoskeletal homeostasis and the lack of known effectors, our research focused on this enzyme. We developed docking and pharmacophore search models for potential αTAT1 modulators using molecular interaction analysis and existing crystal structures. The activity of identified hits was then confirmed via confocal microscopy, allowing for further tuning of our screening models and potential hit-to-lead optimization.