title: Functional reconstitution of Arabidopsis thaliana plant uncoupling mitochondrial protein (AtPUMP1) expressed in Escherichia coli.
abstract: The Arabidopsis thaliana uncoupling protein (UCP) gene was expressed in Escherichia coli and isolated protein reconstituted into liposomes. Linoleic acid-induced H+ fluxes were sensitive to purine nucleotide inhibition with an apparent K(i) (in mM) of 0.8 (GDP), 0.85 (ATP), 0.98 (GTP), and 1.41 (ADP); the inhibition was pH-dependent. Kinetics of AtPUMP1-mediated H+ fluxes were determined for lauric, myristic, palmitic, oleic, linoleic, and linolenic acids. Properties of recombinant AtPUMP1 indicate that it represents a plant counterpart of animal UCP2 or UCP3. This work brings the functional and genetic approaches together for the first time, providing strong support that AtPUMP1 is truly an UCP.
authors.affiliation: Department of Membrane Transport Biophysics, Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
authors.forename: J
authors.initials: J
authors.lastname: Borecký
authors.forename: I G
authors.initials: IG
authors.lastname: Maia
authors.forename: A D
authors.initials: AD
authors.lastname: Costa
authors.forename: P
authors.initials: P
authors.lastname: Jezek
authors.forename: H
authors.initials: H
authors.lastname: Chaimovich
authors.forename: P B
authors.initials: PB
authors.lastname: de Andrade
authors.forename: A E
authors.initials: AE
authors.lastname: Vercesi
authors.forename: P
authors.initials: P
authors.lastname: Arruda
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins; D015153:Blotting, Western; D002352:Carrier Proteins / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D006859:Hydrogen / Q000378:metabolism; D007473:Ion Channels; D007700:Kinetics; D019787:Linoleic Acid / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism*; D026901:Membrane Transport Proteins; D024101:Mitochondrial Proteins; D000071245:Mitochondrial Uncoupling Proteins; D009841:Oligonucleotides / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D000071256:Uncoupling Protein 1; D000071246:Uncoupling Protein 2; D000071247:Uncoupling Protein 3
chemical_list: D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D007473:Ion Channels; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D024101:Mitochondrial Proteins; D000071245:Mitochondrial Uncoupling Proteins; D009841:Oligonucleotides; C000604370:PUMP2 protein, Arabidopsis; D011506:Proteins; D011994:Recombinant Proteins; D000071256:Uncoupling Protein 1; D000071246:Uncoupling Protein 2; D000071247:Uncoupling Protein 3; D006153:Guanosine Diphosphate; D000244:Adenosine Diphosphate; D006859:Hydrogen; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D019787:Linoleic Acid