Cysteine residues are not essential for uncoupling protein function.
The uncoupling protein (UCP) of brown adipose tissue is a regulated proton carrier which allows uncoupling of mitochondrial respiration from ATP synthesis and, therefore, dissipation of metabolic energy as heat. In this article we demonstrate that, when UCP is expressed in Saccharomyces cerevisiae, it retains all its functional properties: proton and chloride transport, high-affinity binding of nucleotides and regulation of proton conductance by nucleotides and fatty acids. Site-directed mutagenesis demonstrates that sequential replacement by serine of cysteine residues in the UCP does not affect either its uncoupling activity or its regulation by nucleotides and fatty acids, and therefore establishes that none of the seven cysteine residues present in the wild-type UCP is critical for its activity. These data indicate that transport models involving essential thiol groups can be discounted and that chemical modification data require critical re-evaluation.
Arechaga
I
I
Centro de Investigaciones Biológicas, C.S.I.C., Madrid, Spain.
Raimbault
S
S
Prieto
S
S
Levi-Meyrueis
C
C
Zaragoza
P
P
Miroux
B
B
Ricquier
D
D
Bouillaud
F
F
Rial
E
E
D001483:Base Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism*; D018076:DNA, Complementary; D006153:Guanosine Diphosphate / Q000378:metabolism; D007473:Ion Channels; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010100:Oxygen / Q000378:metabolism; D010539:Permeability; D011522:Protons; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014475:Uncoupling Agents / Q000737:chemistry / Q000378:metabolism*; D000071256:Uncoupling Protein 1
D002352:Carrier Proteins; D002712:Chlorides; D018076:DNA, Complementary; D007473:Ion Channels; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D011522:Protons; D014475:Uncoupling Agents; D000071256:Uncoupling Protein 1; D006153:Guanosine Diphosphate; D003545:Cysteine; D010100:Oxygen