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Functional reconstitution of Arabidopsis thaliana plant uncoupling mitochondrial protein ( At PUMP1) expressed in Escherichia coli
Author(s) -
Borecký Jirı́,
Maia Ivan G.,
Costa Alexandre D.T.,
Ježek Petr,
Chaimovich Hernan,
de Andrade Paula B.M.,
Vercesi Anı́bal E.,
Arruda Paulo
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)02835-6
Subject(s) - uncoupling protein , biochemistry , escherichia coli , arabidopsis thaliana , biology , linoleic acid , palmitic acid , chemistry , gene , fatty acid , mutant , adipose tissue , brown adipose tissue
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 At PUMP1‐mediated H + fluxes were determined for lauric, myristic, palmitic, oleic, linoleic, and linolenic acids. Properties of recombinant At PUMP1 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 At PUMP1 is truly an UCP.