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Adenosine monophosphate binding stabilizes the KTN domain of the Shewanella denitrificans Kef potassium efflux system

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Christos Pliotas, Samuel C Grayer, Silvia Ekkerman, Anthony K N Chan, Jess Healy, Phedra Marius, Wendy Bartlett, Amjad Khan, Wilian A Cortopassi, Shane A Chandler, Tim Rasmussen, Justin L P Benesch, Robert S Paton, Timothy D W Claridge, Samantha Miller, Ian R Booth, James H Naismith, Stuart J Conway

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Ligand binding is one of the most fundamental properties of proteins. Ligand functions fall into three basic types: substrates, regulatory molecules, and cofactors essential to protein stability, reactivity, or enzyme-substrate complex formation. The regulation of potassium ion movement in bacteria is predominantly under the control of regulatory ligands that gate the relevant channels and transporters, which possess subunits or domains that contain Rossmann folds (RFs). Here we demonstrate that adenosine monophosphate (AMP) is bound to both RFs of the dimeric bacterial Kef potassium efflux system (Kef), where it plays a structural role. We conclude that AMP binds with high affinity, ensuring that the site is fully occupied at all times in the cell. Loss of the ability to bind AMP, we demonstrate, causes protein, and likely dimer, instability and consequent loss of function. Kef system function is regulated via the reversible binding of comparatively low-affinity glutathione-based ligands at the interface between the dimer subunits. We propose this interfacial binding site is itself stabilized, at least in part, by AMP binding.


Original languageEnglish
Pages (from-to)4219-4234
Issue number32
Early online date28 Jun 2017
Publication statusPublished - 15 Aug 2017

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