Theoretical insights on helix repacking as the origin of P-glycoprotein promiscuity

• Type: Article in International Scientific Journal
• Date: 2020

Authors

Bonito, CA (Author) Other; Ferreira, RJ (Author) Other; Ferreira, MJU (Author) Other; Gillet, JP (Author) Other; Natalia N D S Cordeiro (Author) FCUP; dos Santos, DJVA (Author) Other

Journal

Title: Scientific Reports

Vol. 10

ISSN: 2045-2322

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 9 Citations

Scopus - 10 Citations

Other information

Authenticus ID: P-00S-BB4

DOI: 10.1038/s41598-020-66587-5

Abstract (EN): P-glycoprotein (P-gp, ABCB1) overexpression is, currently, one of the most important multidrug resistance (MDR) mechanisms in tumor cells. Thus, modulating drug efflux by P-gp has become one of the most promising approaches to overcome MDR in cancer. Yet, more insights on the molecular basis of drug specificity and efflux-related signal transmission mechanism between the transmembrane domains (TMDs) and the nucleotide binding domains (NBDs) are needed to develop molecules with higher selectivity and efficacy. Starting from a murine P-gp crystallographic structure at the inward-facing conformation (PDB ID: 4Q9H), we evaluated the structural quality of the herein generated human P-gp homology model. This initial human P-gp model, in the presence of the "linker" and inserted in a suitable lipid bilayer, was refined through molecular dynamics simulations and thoroughly validated. The best human P-gp model was further used to study the effect of four single-point mutations located at the TMDs, experimentally related with changes in substrate specificity and drug-stimulated ATPase activity. Remarkably, each P-gp mutation is able to induce transmembrane alpha-helices (TMHs) repacking, affecting the drug-binding pocket volume and the drug-binding sites properties (e.g. volume, shape and polarity) finally compromising drug binding at the substrate binding sites. Furthermore, intracellular coupling helices (ICH) also play an important role since changes in the TMHs rearrangement are shown to have an impact in residue interactions at the ICH-NBD interfaces, suggesting that identified TMHs repacking affect TMD-NBD contacts and interfere with signal transmission from the TMDs to the NBDs.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 13