Identifying New Renal Protein Interactions of Furosemide with a ReverseDock-MD Approach

Background: Furosemide is known to interact with several proteins. A virtual screen of furosemide binding against a library of proteins found in the human kidney has been conducted with the aim of identifying possible interactions. Methods: The initial virtual screen was performed using the automated ReverseDock server. This service obtained the AlphaFold 2 structures of 44 proteins meeting the inclusion criteria and provided docking results obtained using AutoDock Vina. The stability of the best docked poses was challenged by three 10 ns molecular dynamics (MD) simulations run using GROMACS-on-Colab. Results: Through analysis of the ligand-protein pairings obtained from ReverseDock and based on a binding location with possible functional consequences, 12 protein models were selected for assessment using MD. Stable ligand behaviour was observed for 8 proteins including plausible novel interactions with Sodium/mannose cotransporter (SGLT5), Sodium/bile acid cotransporter 7, Glucose-6-phosphate exchanger. Interactions with proteins known to interact with furosemide including aquaporin 1, organic anion transporter 1 (OAT1) and organic anion transporter 3 (OAT3) were also observed. Conclusion: Plausible novel interactions between furosemide and sodium/mannose cotransporter, Glucose-6-phosphate exchanger, and sodium/bile transporter 7 have been identified in this study. Stable interactions in silico were identified for OAT1 and OAT3; proteins known to transport furosemide.A selection of furosemide docking results from this study