Rhodopsins, most of which are proton pumps generating transmembrane electrochemical proton gradients, span all three domains of life, are abundant in the biosphere, and could play a crucial role in the early evolution of life on earth. Whereas archaeal and bacterial proton pumps are among the best structurally characterized proteins, rhodopsins from unicellular eukaryotes have not been well characterized. To fill this gap in the current understanding of the proton pumps and to gain insight into the evolution of rhodopsins using a structure-based approach, we performed a structural and functional analysis of the light-driven proton pump LR (Mac) from the pathogenic fungus Leptosphaeria maculans. The first high-resolution structure of fungi rhodopsin and its functional properties reveal the striking similarity of its membrane part to archaeal but not to bacterial rhodopsins. We show that an unusually long N-terminal region stabilizes the protein through direct interaction with its extracellular loop (ECL2). We compare to our knowledge all available structures and sequences of outward light-driven proton pumps and show that eukaryotic and archaeal proton pumps, most likely, share a common ancestor.

ELI Beamlines Institute of Physics Czech Academy of Sciences Prague Czech Republic

European Molecular Biology Laboratory Hamburg unit c o DESY Hamburg Germany

European Molecular Biology Laboratory Hamburg Unit Notkestrasse 25a Hamburg Germany

Institut de Biologie Structurale Université Grenoble Alpes CEA CNRS Grenoble France

Institut für Biologische Informationsprozesse Molekular und Zellphysiologie Forschungszentrum Jülich Jülich Germany

Institute for Biophysical Chemistry Hannover Medical School Hannover Germany

Institute for Safety Problems of Nuclear Power Plants NAS of Ukraine Kyiv Ukraine

Institute of Biological Information Processing ForschungszentrumJülich Jülich Germany

Institute of Crystallography University of Aachen Aachen Germany

Joint Institute for Nuclear Research Dubna Russia

JuStruct Jülich Center for Structural Biology Forschungszentrum Jülich Jülich Germany

Max Planck Institute of Biophysics Frankfurt am Main Germany

National Center for Biotechnology Information National Library of Medicine National Institutesof Health Bethesda MD United States

Research Center for Molecular Mechanisms of Aging and Age related Diseases Moscow Institute of Physics and Technology Dolgoprudny Russia

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