title: The Peroxisomal NAD Carrier from Arabidopsis Imports NAD in Exchange with AMP.
abstract: Cofactors such as NAD, AMP, and Coenzyme A (CoA) are essential for a diverse set of reactions and pathways in the cell. Specific carrier proteins are required to distribute these cofactors to different cell compartments, including peroxisomes. We previously identified a peroxisomal transport protein in Arabidopsis (Arabidopsis thaliana) called the peroxisomal NAD carrier (PXN). When assayed in vitro, this carrier exhibits versatile transport functions, e.g. catalyzing the import of NAD or CoA, the exchange of NAD/NADH, and the export of CoA. These observations raise the question about the physiological function of PXN in plants. Here, we used Saccharomyces cerevisiae to address this question. First, we confirmed that PXN, when expressed in yeast, is active and targeted to yeast peroxisomes. Secondl, detailed uptake analyses revealed that the CoA transport function of PXN can be excluded under physiological conditions due to its low affinity for this substrate. Third, we expressed PXN in diverse mutant yeast strains and investigated the suppression of the mutant phenotypes. These studies provided strong evidences that PXN was not able to function as a CoA transporter or a redox shuttle by mediating a NAD/NADH exchange, but instead catalyzed the import of NAD into peroxisomes against AMP in intact yeast cells.
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Carlo W T
authors.identifier: 0000-0001-5182-8076
authors.initials: CW
authors.lastname: van Roermund
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Martin G
authors.initials: MG
authors.lastname: Schroers
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Jan
authors.initials: J
authors.lastname: Wiese
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Fabio
authors.initials: F
authors.lastname: Facchinelli
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Samantha
authors.identifier: 0000-0002-6853-1399
authors.initials: S
authors.lastname: Kurz
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Sabrina
authors.initials: S
authors.lastname: Wilkinson
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Lennart
authors.initials: L
authors.lastname: Charton
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Ronald J A
authors.initials: RJ
authors.lastname: Wanders
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Hans R
authors.initials: HR
authors.lastname: Waterham
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.).
authors.forename: Andreas P M
authors.identifier: 0000-0003-0970-4672
authors.initials: AP
authors.lastname: Weber
authors.affiliation: Laboratory Genetic Metabolic Diseases, Laboratory Division, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands (C.W.T.v.R., R.J.A.W., H.R.W.); andInstitute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University, 40225 Düsseldorf, Germany (M.G.S., J.W., F.F., S.K., S.W., L.C., A.P.M.W., N.L.) nicole.linka@hhu.de.
authors.forename: Nicole
authors.identifier: 0000-0002-1400-717X
authors.initials: N
authors.lastname: Link
mesh_terms: D000249:Adenosine Monophosphate / Q000378:metabolism*; D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D003065:Coenzyme A / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D033681:Mitochondrial Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D024101:Mitochondrial Proteins; D009243:NAD / Q000378:metabolism*; D033704:Nucleotide Transport Proteins; D027701:Organic Cation Transport Proteins / Q000235:genetics; D020675:Peroxisomes / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D017384:Sequence Deletion
chemical_list: D029681:Arabidopsis Proteins; D033681:Mitochondrial Membrane Transport Proteins; D024101:Mitochondrial Proteins; C508687:Ndt1 protein, S cerevisiae; D033704:Nucleotide Transport Proteins; D027701:Organic Cation Transport Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C574424:solute carrier family 25 member 17 protein, Arabidopsis; D009243:NAD; D000249:Adenosine Monophosphate; D008291:Malate Dehydrogenase; D003065:Coenzyme A