Proton transfer activity of the reconstituted Mycobacterium tuberculosis MmpL3 is modulated by substrate mimics and inhibitors

Transporters belonging to the Resistance-Nodulation-cell Division (RND) superfamily of proteins such as MmpL3 and its analogs are the focus of intense investigations due to their importance in the physiology of species and antimycobacterial drug discovery. These transporters deliver trehalose monomy...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2022-07, Vol.119 (30), p.e2113963119
Main Authors: Stevens, Casey M, Babii, Svitlana O, Pandya, Amitkumar N, Li, Wei, Li, Yupeng, Mehla, Jitender, Scott, Robyn, Hegde, Pooja, Prathipati, Pavan K, Acharya, Atanu, Liu, Jinchan, Gumbart, James C, North, Jeffrey, Jackson, Mary, Zgurskaya, Helen I
Format: Article
Language:English
Subjects:
Bacterial Proteins - antagonists & inhibitors
Bacterial Proteins - chemistry
Biological Sciences
Cell division
Corynebacterium
Inhibitors
Ion Transport
Lipid Bilayers - chemistry
Lipids
Membrane Transport Proteins - chemistry
Membranes
Molecular dynamics
Mycobacterium tuberculosis
Mycolic Acids - metabolism
Nodulation
Periplasm
pH effects
Phospholipids
Proteins
Protonmotive force
Protons
Substrate inhibition
Substrate Specificity
Substrates
Translocation
Trehalose
Tuberculosis
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Transporters belonging to the Resistance-Nodulation-cell Division (RND) superfamily of proteins such as MmpL3 and its analogs are the focus of intense investigations due to their importance in the physiology of species and antimycobacterial drug discovery. These transporters deliver trehalose monomycolates, the precursors of major lipids of the outer membrane, to the periplasm by a proton motive force-dependent mechanism. In this study, we successfully purified, from native membranes, the full-length and the C-terminal truncated MmpL3 and CmpL1 proteins and reconstituted them into proteoliposomes. We also generated a series of substrate mimics and inhibitors specific to these transporters, analyzed their activities in the reconstituted proteoliposomes, and carried out molecular dynamics simulations of the model MmpL3 transporter at different pH. We found that all reconstituted proteins facilitate proton translocation across a phospholipid bilayer, but MmpL3 and CmpL1 differ dramatically in their responses to pH and interactions with substrate mimics and indole-2-carboxamide inhibitors. Our results further suggest that some inhibitors abolish the transport activity of MmpL3 and CmpL1 by inhibition of proton translocation.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.2113963119