Deciphering the conformational transitions of LIMK2 active and inactive states to ponder specific druggable states through microsecond scale molecular dynamics simulation.

LIMK2 inhibitors are one of the potential therapeutic modalities for treating various diseases. In the current scenario, there is a paucity of effective LIMK inhibitors that are highly specific with minimal off-target effects. To date, the conformational transitions of LIMK2 from DFGinαCin (CIDI) (active) to DFGoutαCout (CODO) (inactive) states are yet to be probed and are essential for capturing the unique, druggable conformations. Therefore, this study was intended to capture the diverse conformational states of LIMK2 for accelerating the rational identification of conformation specific inhibitors through high-end structural bioinformatics protocols. Hence, in this study, molecular modelling followed by an extensive microsecond timescale of molecular dynamics simulation was performed encompassing perturbation response scanning, metapath, and community analysis towards the conformational sampling of LIMK2. Overall this study precisely identifies the conformational ensemble of LIMK2 the intermediate inactive states namely, CIDO, CinterDinter, CIDinter, CinterDI, CinterDO, CODI, CODinter apart from CIDI and CODO. This also facilitated observing that β8 preceding XDFG, αC (F373, L374), and αD (L413) as the major effectors that may facilitate the regulation of varying conformational transitions among the states. Additionally, the conserved β sheets and the loops namely, C.l, b.l, and G/P.loop were observed to be involved in the metapath for allosteric communication among the intermediates with CIDI and CODO state. Moreover, only the CODO state was observed to have closed type A.l, while the CIDI and other intermediate states except for CIDO were observed to have open-DFG out type A.l, thereby enabling the binding of substrate. Apart from these, the druggable site analysis inferred that the CIDI and CODO states harbor prominent druggable sites spanning the conserved N-lobe, while the intermediates were observed to have unraveled allosteric druggable sites distal from the ATP binding site, majorly spanning the C-lobe of LIMK2. Thus, this study provides potential insights into the intermediate conformational druggable states of LIMK2 and also the druggable conformations, especially the inactive states of LIMK2, as a specific therapeutic targeting mode. Thus, providing a widened avenue to ponder the allosteric sites or the isoform selectivity conformations for targeting LIMK2 in various disease conditions.