Engineered MATE multidrug transporters reveal two functionally distinct ion-coupling pathways in NorM from Vibrio cholerae

Abstract Multidrug and toxic compound extrusion (MATE) transport proteins confer multidrug resistance on pathogenic microorganisms affect pharmacokinetics in mammals. Our understanding of how MATE transporters work, has mostly relied protein structures MD simulations. However, the energetics drug not been studied detail. Many utilise electrochemical H + or Na gradient to drive substrate efflux, but NorM-VC from Vibrio cholerae can both forms metabolic energy. To dissect localisation organisation translocation pathways we engineered chimaeric which N-lobe -coupled NorM-PS Pseudomonas stutzeri is fused C-lobe NorM-VC, vice versa. findings binding experiments with chimaeric, mutant wildtype highlight versatile nature energy coupling enables adaptation fluctuating salinity levels natural habitat V. .