1. Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae
- Author
-
Asha V. Nair, Boyan Bai, Himansha Singh, Hendrik W. van Veen, Keiko Shinoda, Hideaki Fujitani, Sagar Raturi, Satoshi Murakami, Raturi, Sagar [0000-0003-4040-7799], Nair, Asha V. [0000-0002-2037-1138], Murakami, Satoshi [0000-0001-5553-7663], van Veen, Hendrik W. [0000-0002-9658-8077], Apollo - University of Cambridge Repository, Nair, Asha V [0000-0002-2037-1138], and van Veen, Hendrik W [0000-0002-9658-8077]
- Subjects
Organic Cation Transport Proteins ,QH301-705.5 ,education ,Mutant ,Medicine (miscellaneous) ,96/34 ,Plasma protein binding ,Antimicrobial resistance ,medicine.disease_cause ,631/45/173 ,38/70 ,Antiporters ,General Biochemistry, Genetics and Molecular Biology ,82/80 ,38/1 ,03 medical and health sciences ,0302 clinical medicine ,Protein structure ,Bacterial Proteins ,medicine ,Biology (General) ,Binding site ,82/83 ,Vibrio cholerae ,030304 developmental biology ,Ions ,0303 health sciences ,Binding Sites ,Chemistry ,Sodium ,article ,Biological Transport ,Transporter ,humanities ,Drug Resistance, Multiple ,Cell biology ,Transport protein ,Enzyme mechanisms ,631/326/22/1434 ,behavior and behavior mechanisms ,Efflux ,General Agricultural and Biological Sciences ,030217 neurology & neurosurgery ,Hydrogen ,Protein Binding - Abstract
Funder: Cambridge Commonwealth, European and International Trust (Cambridge Commonwealth, European & International Trust); doi: https://doi.org/10.13039/501100003343, Funder: The Nehru Trust for Cambridge University, Funder: H.F. and K.S. were supported in part by the Innovative Drug Discovery Infrastructure through the Functional Control of Biomolecular Systems Priority Issue1 in Post-K Supercomputer Development from MEXT (Project ID: hp190171, hp170255, hp180191, and hp150269. K.S. was also supported in part by JSPS KAKENHI Grant Number JP20H05453., Funder: H.F. and K.S. were supported in part by the Innovative Drug Discovery Infrastructure through the Functional Control of Biomolecular Systems Priority Issue1 in Post-K Supercomputer Development from MEXT (Project ID: hp190171, hp170255, hp180191, and hp150269)., Multidrug and toxic compound extrusion (MATE) transport proteins confer multidrug resistance on pathogenic microorganisms and affect pharmacokinetics in mammals. Our understanding of how MATE transporters work, has mostly relied on protein structures and MD simulations. However, the energetics of drug transport has not been studied in detail. Many MATE transporters utilise the electrochemical H+ or Na+ gradient to drive substrate efflux, but NorM-VC from Vibrio cholerae can utilise both forms of metabolic energy. To dissect the localisation and organisation of H+ and Na+ translocation pathways in NorM-VC we engineered chimaeric proteins in which the N-lobe of H+-coupled NorM-PS from Pseudomonas stutzeri is fused to the C-lobe of NorM-VC, and vice versa. Our findings in drug binding and transport experiments with chimaeric, mutant and wildtype transporters highlight the versatile nature of energy coupling in NorM-VC, which enables adaptation to fluctuating salinity levels in the natural habitat of V. cholerae.
- Published
- 2021