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PMID: 11171965 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Uncoupling proteins 2 and 3 are highly active H(+) transporters and highly nucleotide sensitive when activated by coenzyme Q (ubiquinone).

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 98 ·No. 4 ·2001-02-13 ·Pages 1416-21

Echtay KS, Winkler E, Frischmuth K, Klingenberg M

Abstract

Based on the discovery of coenzyme Q (CoQ) as an obligatory cofactor for H(+) transport by uncoupling protein 1 (UCP1) [Echtay, K. S., Winkler, E. & Klingenberg, M. (2000) Nature (London) 408, 609-613] we show here that UCP2 and UCP3 are also highly active H(+) transporters and require CoQ and fatty acid for H(+) transport, which is inhibited by low concentrations of nucleotides. CoQ is proposed to facilitate injection of H(+) from fatty acid into UCP. Human UCP2 and 3 expressed in Escherichia coli inclusion bodies are solubilized, and by exchange of sarcosyl against digitonin, nucleotide binding as measured with 2'-O-[5-(dimethylamino)naphthalene-1-sulfonyl]-GTP can be restored. After reconstitution into vesicles, Cl(-) but no H(+) are transported. The addition of CoQ initiates H(+) transport in conjunction with fatty acids. This increase is fully sensitive to nucleotides. The rates are as high as with reconstituted UCP1 from mitochondria. Maximum activity is at a molar ratio of 1:300 of CoQ:phospholipid. In UCP2 as in UCP1, ATP is a stronger inhibitor than ADP, but in UCP3 ADP inhibits more strongly than ATP. Thus UCP2 and UCP3 are regulated differently by nucleotides, in line with their different physiological contexts. These results confirm the regulation of UCP2 and UCP3 by the same factors CoQ, fatty acids, and nucleotides as UCP1. They supersede reports that UCP2 and UCP3 may not be H(+) transporters.

MeSH Terms
Animals Benzoquinones/metabolism Carrier Proteins/metabolism Coenzymes Cricetinae Dose-Response Relationship, Drug Escherichia coli Fatty Acids/metabolism Humans Hydrogen/metabolism Inclusion Bodies Ion Channels Membrane Transport Proteins Mitochondrial Proteins Nucleotides Proteins/metabolism Structure-Activity Relationship Ubiquinone/analogs & derivatives,metabolism Uncoupling Protein 2 Uncoupling Protein 3
Chemicals
Benzoquinones Carrier Proteins Coenzymes Fatty Acids Ion Channels Membrane Transport Proteins Mitochondrial Proteins Nucleotides Proteins UCP2 protein, human UCP3 protein, human Uncoupling Protein 2 Uncoupling Protein 3 ubiquinone 6 Ubiquinone ubiquinone-O Hydrogen coenzyme Q10 Ubiquinone Q2 Ubiquinone Q1
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Echtay K S
Institute for Physiological Chemistry, University of Munich, Schillerstrasse 44, D-80336 Munich, Germany.
Winkler E
Frischmuth K
Klingenberg M
References (50)
50 references, click to expand
  1. Uncoupling protein-3 is a mediator of thermogenesis regulated by thyroid hormone, beta3-adrenergic agonists, and leptin.
    J Biol Chem. 1997 Sep 26;272(39):24129-32 PMID: 9305858
  2. Thermogenic mechanisms in brown fat.
    Physiol Rev. 1984 Jan;64(1):1-64 PMID: 6320232
  3. Diet-induced changes in uncoupling proteins in obesity-prone and obesity-resistant strains of mice.
    Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):4061-5 PMID: 9520493
  4. A high diffusion coefficient for coenzyme Q10 might be related to a folded structure.
    FEBS Lett. 1998 Apr 10;426(1):77-80 PMID: 9598982
  5. The uncoupling proteins, a review.
    Eur J Endocrinol. 1998 Jul;139(1):1-9 PMID: 9703368
  6. A ubiquinone-binding site regulates the mitochondrial permeability transition pore.
    J Biol Chem. 1998 Oct 2;273(40):25734-40 PMID: 9748242
  7. Thermogenic responses in brown fat cells are fully UCP1-dependent. UCP2 or UCP3 do not substitute for UCP1 in adrenergically or fatty scid-induced thermogenesis.
    J Biol Chem. 2000 Aug 18;275(33):25073-81 PMID: 10825155
  8. The mitochondrial permeability transition: its molecular mechanism and role in reperfusion injury.
    Biochem Soc Symp. 1999;66:181-203 PMID: 10989667
  9. Three classes of ubiquinone analogs regulate the mitochondrial permeability transition pore through a common site.
    J Biol Chem. 2000 Sep 22;275(38):29521-7 PMID: 10889201
  10. The bulk of UCP3 expressed in yeast cells is incompetent for a nucleotide regulated H+ transport.
    FEBS Lett. 2000 Sep 1;480(2-3):265-70 PMID: 11034342
  11. Coenzyme Q is an obligatory cofactor for uncoupling protein function.
    Nature. 2000 Nov 30;408(6812):609-13 PMID: 11117751
  12. Genomic organization and regulation by dietary fat of the uncoupling protein 3 and 2 genes.
    Biochem Biophys Res Commun. 1999 Mar 5;256(1):27-32 PMID: 10066417
  13. Recombinant human uncoupling protein-3 increases thermogenesis in yeast cells.
    FEBS Lett. 1999 Apr 1;448(1):57-61 PMID: 10217410
  14. Assessment of uncoupling activity of uncoupling protein 3 using a yeast heterologous expression system.
    FEBS Lett. 1999 Apr 23;449(2-3):129-34 PMID: 10338118
  15. Regulation of UCP3 by nucleotides is different from regulation of UCP1.
    FEBS Lett. 1999 Apr 30;450(1-2):8-12 PMID: 10350047
  16. Contributions of studies on uncoupling proteins to research on metabolic diseases.
    J Intern Med. 1999 Jun;245(6):637-42 PMID: 10395193
  17. Assessment of uncoupling activity of the human uncoupling protein 3 short form and three mutants of the uncoupling protein gene using a yeast heterologous expression system.
    FEBS Lett. 1999 Jul 9;454(3):201-6 PMID: 10431807
  18. Thyroid hormone and other regulators of uncoupling proteins.
    Int J Obes Relat Metab Disord. 1999 Jun;23 Suppl 6:S56-9 PMID: 10454126
  19. Transport function and regulation of mitochondrial uncoupling proteins 2 and 3.
    J Biol Chem. 1999 Sep 10;274(37):26003-7 PMID: 10473545
  20. Difference in induction of uncoupling protein genes in adipose tissues between young and old rats during cold exposure.
    FEBS Lett. 1999 Sep 17;458(2):157-61 PMID: 10481056
  21. The mitochondrial uncoupling protein-2: current status.
    Int J Biochem Cell Biol. 1999 Nov;31(11):1261-78 PMID: 10605819
  22. UCP2 and UCP3 rise in starved rat skeletal muscle but mitochondrial proton conductance is unchanged.
    FEBS Lett. 1999 Dec 3;462(3):257-60 PMID: 10622707
  23. Properties of the human long and short isoforms of the uncoupling protein-3 expressed in yeast cells.
    FEBS Lett. 1999 Dec 3;462(3):411-5 PMID: 10622736
  24. The uncoupling protein homologues: UCP1, UCP2, UCP3, StUCP and AtUCP.
    Biochem J. 2000 Jan 15;345 Pt 2:161-79 PMID: 10620491
  25. Uncoupling protein 3: its possible biological role and mode of regulation in rodents and humans.
    J Bioenerg Biomembr. 1999 Oct;31(5):467-73 PMID: 10653475
  26. UCP1: the original uncoupling protein--and perhaps the only one? New perspectives on UCP1, UCP2, and UCP3 in the light of the bioenergetics of the UCP1-ablated mice.
    J Bioenerg Biomembr. 1999 Oct;31(5):475-91 PMID: 10653476
  27. Isolation of the uncoupling protein from brown adipose tissue mitochondria.
    FEBS Lett. 1980 May 5;113(2):299-303 PMID: 7389900
  28. Mice overexpressing human uncoupling protein-3 in skeletal muscle are hyperphagic and lean.
    Nature. 2000 Jul 27;406(6794):415-8 PMID: 10935638
  29. The biochemistry of an inefficient tissue: brown adipose tissue.
    Essays Biochem. 1985;20:110-64 PMID: 3928372
  30. Mechanism and evolution of the uncoupling protein of brown adipose tissue.
    Trends Biochem Sci. 1990 Mar;15(3):108-12 PMID: 2158156
  31. Functional reconstitution of rat uncoupling protein following its high level expression in yeast.
    J Biol Chem. 1991 Jun 25;266(18):11871-5 PMID: 2050682
  32. Functional expression of the rat brown adipose tissue uncoupling protein in Saccharomyces cerevisiae.
    Mol Microbiol. 1992 Feb;6(3):363-70 PMID: 1552849
  33. Cysteine residues are not essential for uncoupling protein function.
    Biochem J. 1993 Dec 15;296 ( Pt 3):693-700 PMID: 8280067
  34. Effect of fatty acids on H+ transport activity of the reconstituted uncoupling protein.
    J Biol Chem. 1994 Jan 28;269(4):2508-15 PMID: 8300577
  35. Fluorescent nucleotide derivatives as specific probes for the uncoupling protein: thermodynamics and kinetics of binding and the control by pH.
    Biochemistry. 1995 Jan 10;34(1):349-60 PMID: 7819218
  36. Mitochondrial ADP/ATP carrier can be reversibly converted into a large channel by Ca2+.
    Biochemistry. 1996 Jul 2;35(26):8483-8 PMID: 8679608
  37. Identification of the pH sensor for nucleotide binding in the uncoupling protein from brown adipose tissue.
    Biochemistry. 1997 Jan 7;36(1):148-55 PMID: 8993328
  38. Uncoupling protein-2: a novel gene linked to obesity and hyperinsulinemia.
    Nat Genet. 1997 Mar;15(3):269-72 PMID: 9054939
  39. Cloning and characterization of an uncoupling protein homolog: a potential molecular mediator of human thermogenesis.
    Diabetes. 1997 May;46(5):900-6 PMID: 9133562
  40. Uncoupling protein-3: a new member of the mitochondrial carrier family with tissue-specific expression.
    FEBS Lett. 1997 May 12;408(1):39-42 PMID: 9180264
  41. UCP3: an uncoupling protein homologue expressed preferentially and abundantly in skeletal muscle and brown adipose tissue.
    Biochem Biophys Res Commun. 1997 Jun 9;235(1):79-82 PMID: 9196039
  42. Mutagenesis of the uncoupling protein of brown adipose tissue. Neutralization Of E190 largely abolishes pH control of nucleotide binding.
    Biochemistry. 1997 Jul 8;36(27):8253-60 PMID: 9204870
  43. Effects of fatty acids on uncoupling protein-2 expression in the rat heart.
    FASEB J. 2000 Mar;14(3):495-502 PMID: 10698964
  44. Site-directed mutagenesis identifies residues in uncoupling protein (UCP1) involved in three different functions.
    Biochemistry. 2000 Mar 28;39(12):3311-7 PMID: 10727223
  45. Downregulation of skeletal muscle UCP-3 gene expression during refeeding is prevented by cold exposure.
    Pflugers Arch. 2000 Apr;439(6):723-9 PMID: 10784346
  46. Uncoupling proteins 1 and 3 are regulated differently.
    Biochemistry. 2000 May 16;39(19):5845-51 PMID: 10801335
  47. Lack of obesity and normal response to fasting and thyroid hormone in mice lacking uncoupling protein-3.
    J Biol Chem. 2000 May 26;275(21):16251-7 PMID: 10748195
  48. Energy metabolism in uncoupling protein 3 gene knockout mice.
    J Biol Chem. 2000 May 26;275(21):16258-66 PMID: 10748196
  49. Uncoupling proteins 2 and 3: potential regulators of mitochondrial energy metabolism.
    Diabetes. 2000 Feb;49(2):143-56 PMID: 10868929
  50. Uncoupling: new approaches to an old problem of bioenergetics.
    Biochim Biophys Acta. 1998 Feb 25;1363(2):100-24 PMID: 9507078
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.98.4.1416
Published
2001-02-13
Pages
1416-21
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC29271
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.98.4.1416 PMC Full Text View journal details
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