title: Origin and evolution of transporter substrate specificity within the NPF family.
abstract: Despite vast diversity in metabolites and the matching substrate specificity of their transporters, little is known about how evolution of transporter substrate specificities is linked to emergence of substrates via evolution of biosynthetic pathways. Transporter specificity towards the recently evolved glucosinolates characteristic ofBrassicalesis shown to evolve prior to emergence of glucosinolate biosynthesis. Furthermore, we show that glucosinolate transporters belonging to the ubiquitous NRT1/PTR FAMILY (NPF) likely evolved from transporters of the ancestral cyanogenic glucosides found across more than 2500 species outside of theBrassicales. Biochemical characterization of orthologs along the phylogenetic lineage from cassava toA. thaliana,suggests that alterations in the electrogenicity of the transporters accompanied changes in substrate specificity. Linking the evolutionary path of transporter substrate specificities to that of the biosynthetic pathways, exemplify how transporter substrate specificities originate and evolve as new biosynthesis pathways emerge.
authors.affiliation: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.|Copenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.
authors.forename: Morten Egevang
authors.identifier: http://orcid.org/0000-0001-6503-0495
authors.initials: ME
authors.lastname: Jørgensen
authors.affiliation: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.|Copenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.
authors.forename: Deyang
authors.identifier: http://orcid.org/0000-0003-1015-6458
authors.initials: D
authors.lastname: Xu
authors.affiliation: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.|Copenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.
authors.forename: Christoph
authors.initials: C
authors.lastname: Crocoll
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Heidi Asschenfeldt
authors.initials: HA
authors.lastname: Ernst
authors.affiliation: Centro de Bioinformática y Simulación Molecular (CBSM), Universidad de Talca, Talca, Chile.|Instituto de Innovación Basada en Ciencia, Universidad de Talca, Talca, Chile.
authors.forename: David
authors.initials: D
authors.lastname: Ramírez
authors.affiliation: Center for Plant Plasticity, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.
authors.forename: Mohammed Saddik
authors.initials: MS
authors.lastname: Motawia
authors.affiliation: Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.
authors.forename: Carl Erik
authors.initials: CE
authors.lastname: Olsen
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Osman
authors.initials: O
authors.lastname: Mirza
authors.affiliation: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.|Copenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.
authors.forename: Hussam Hassan
authors.identifier: http://orcid.org/0000-0001-6660-0509
authors.initials: HH
authors.lastname: Nour-Eldin
authors.affiliation: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.|Copenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.
authors.forename: Barbara Ann
authors.identifier: http://orcid.org/0000-0002-3742-702X
authors.initials: BA
authors.lastname: Halkier
mesh_terms: D019143:Evolution, Molecular; D005961:Glucosinolates / Q000378:metabolism*; D019684:Magnoliopsida / Q000235:genetics* / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D013379:Substrate Specificity
chemical_list: D005961:Glucosinolates; D026901:Membrane Transport Proteins
keywords: A. thaliana; biochemistry; cyanogenic glucoside transport; indole glucosinolate transport; metabolite transporters; plant biology; substrate specificity and electrogenicity; transporter evolution; xenopus