Generation of an electrochemical proton gradient is the first step of cell bioenergetics. In prokaryotes, the gradient is created by outward membrane protein proton pumps. Inward plasma membrane native proton pumps are yet unknown. We describe comprehensive functional studies of the representatives of the yet noncharacterized xenorhodopsins from Nanohaloarchaea family of microbial rhodopsins. They are inward proton pumps as we demonstrate in model membrane systems, Escherichia coli cells, human embryonic kidney cells, neuroblastoma cells, and rat hippocampal neuronal cells. We also solved the structure of a xenorhodopsin from the nanohalosarchaeon Nanosalina (NsXeR) and suggest a mechanism of inward proton pumping. We demonstrate that the NsXeR is a powerful pump, which is able to elicit action potentials in rat hippocampal neuronal cells up to their maximal intrinsic firing frequency. Hence, inwardly directed proton pumps are suitable for light-induced remote control of neurons, and they are an alternative to the well-known cation-selective channelrhodopsins.

Division for Structural Biochemistry Hannover Medical School Hannover Germany

European Synchrotron Radiation Facility 38027 Grenoble France

Evolutionary Genomics Group Departamento de Producción Vegetal y Microbiología Universidad Miguel Hernández San Juan de Alicante Alicante Spain

Friedrich Miescher Institute for Biomedical Research Basel Switzerland

Institut de Biologie Structurale Jean Pierre Ebel Université Grenoble Alpes Commissariat à l'Energie Atomique et aux Energies Alternatives CNRS Grenoble France

Institute for Biodiversity and Ecosystem Dynamics University of Amsterdam Amsterdam Netherlands

Institute for Biophysical Chemistry Hannover Medical School Hannover Germany

Institute of Complex Systems ICS 6 Structural Biochemistry Research Centre Jülich Jülich Germany

Institute of Crystallography RWTH Aachen University Aachen Germany

Institute of Hydrobiology Department of Aquatic Microbial Ecology Biology Centre of the Czech Academy of Sciences České Budějovice Czech Republic

Joint Institute for Nuclear Research Dubna Russia

Max Planck Institute of Biophysics Frankfurt am Main Germany

Moscow Institute of Physics and Technology Dolgoprudny Russia

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Schizorhodopsins: A family of rhodopsins from Asgard archaea that function as light-driven inward H+ pumps