Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1983 Mar 15;210(3):859–866. doi: 10.1042/bj2100859

An immunological study of the uncoupling protein of brown adipose tissue mitochondria.

D Ricquier, J P Barlet, J M Garel, M Combes-George, M P Dubois
PMCID: PMC1154300  PMID: 6409080

Abstract

1. Ewes were injected with purified 32,000-Mr uncoupling protein from mitochondria of brown adipose tissue of cold-adapted rats in order to raise antibodies. 2. The existence of antibodies in the plasma of ewes and the cross-reactivity of plasmas were demonstrated and studied by 125I-labelled antigen-antibody reaction, double immunodiffusion, the inhibition of GDP binding to the 32,000 Mr protein and by immunohistochemistry. 3. The antibodies raised against the homogeneous protein yielded a single immunoprecipitation band with detergent-solubilized mitochondrial membranes of brown adipose tissue from rat, hamster, guinea-pig, rabbit and with the purified uncoupling protein of these animals. No immunoprecipitation was obtained with the protein purified from brown adipose tissue of term lamb foetus. 4. The GDP-binding activity of the uncoupling protein (isolated or in solubilized membranes) was largely inhibited by the antiserum. 5. The anti-(rat uncoupling protein) could not cross-react with solubilized membranes from liver or muscle, nor with the purified beef heart or rat liver ADP/ATP translocator.

Full text

PDF
860

Images in this article

861Fig. 1.
on p.861
862Fig. 3.
on p.862
864Fig. 4.
on p.864

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cabral F., Schatz G. High resolution one- and two- dimensional electrophoretic analysis of mitochondrial membrane polypeptides. Methods Enzymol. 1979;56:602–613. doi: 10.1016/0076-6879(79)56057-1. [DOI] [PubMed] [Google Scholar]
  2. Desautels M., Himms-Hagen J. Roles of noradrenaline and protein synthesis in the cold-induced increase in purine nucleotide binding by rat brown adipose tissue mitochondria. Can J Biochem. 1979 Jun;57(6):968–976. doi: 10.1139/o79-118. [DOI] [PubMed] [Google Scholar]
  3. Eiermann W., Aquila H., Klingenberg M. Immunological characterization of the ADP, ATP translocator protein isolated from mitochondria of liver, heart and other organs. Evidence for an organ specificity. FEBS Lett. 1977 Mar 1;74(2):209–214. doi: 10.1016/0014-5793(77)80848-x. [DOI] [PubMed] [Google Scholar]
  4. Foxwell B. M., Tanner M. J. Synthesis of the erythrocyte anion-transport protein. Immunochemical study of its incorporation into the plasma membrane of erythroid cells. Biochem J. 1981 Apr 1;195(1):129–137. doi: 10.1042/bj1950129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Garel J. M., Care A. D., Barlet J. P. A radioimmunoassay for ovine calcitonin: an evaluation of calcitonin secretion during gestation, lactation and foetal life. J Endocrinol. 1974 Sep;62(3):497–509. doi: 10.1677/joe.0.0620497. [DOI] [PubMed] [Google Scholar]
  6. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  7. Klingenberg M., Aquila H., Riccio P. Isolation of functional membrane proteins related to or identical with the ADP, ATP carrier of mitochondria. Methods Enzymol. 1979;56:407–414. doi: 10.1016/0076-6879(79)56039-x. [DOI] [PubMed] [Google Scholar]
  8. Lin C. S., Klingenberg M. Isolation of the uncoupling protein from brown adipose tissue mitochondria. FEBS Lett. 1980 May 5;113(2):299–303. doi: 10.1016/0014-5793(80)80613-2. [DOI] [PubMed] [Google Scholar]
  9. Locke R. M., Nicholls D. G. A re-evaluation of the role of fatty acids in the physiological regulation of the proton conductance of brown adipose tissue mitochondria. FEBS Lett. 1981 Dec 7;135(2):249–252. doi: 10.1016/0014-5793(81)80793-4. [DOI] [PubMed] [Google Scholar]
  10. Nedergaard J., Cannon B. Overview--preparation and properties of mitochondria from different sources. Methods Enzymol. 1979;55:3–28. doi: 10.1016/0076-6879(79)55003-4. [DOI] [PubMed] [Google Scholar]
  11. Nicholls D. G. Brown adipose tissue mitochondria. Biochim Biophys Acta. 1979 Jul 3;549(1):1–29. doi: 10.1016/0304-4173(79)90016-8. [DOI] [PubMed] [Google Scholar]
  12. Rechler M. M., Fryklund L., Nissley S., Hall K., Podskalny J. M., Skottner A., Moses A. C. Purified human somatomedin A and rat multiplication stimulating activity. Mitogens for cultured fibroblasts that cross-react with the same growth peptide receptors. Eur J Biochem. 1978 Jan 2;82(1):5–12. doi: 10.1111/j.1432-1033.1978.tb11991.x. [DOI] [PubMed] [Google Scholar]
  13. Ricquier D., Gervais C., Kader J. C., Hemon P. Partial purification by guanosine-5'-diphosphate--agarose affinity chromatography of the 32,000 molecular weight polypeptide from mitochondria of brown adipose tissue. FEBS Lett. 1979 May 1;101(1):35–38. [PubMed] [Google Scholar]
  14. Ricquier D., Kader J. C. Mitochondrial protein alteration in active brown fat: a soidum dodecyl sulfate-polyacrylamide gel electrophoretic study. Biochem Biophys Res Commun. 1976 Dec 6;73(3):577–583. doi: 10.1016/0006-291x(76)90849-4. [DOI] [PubMed] [Google Scholar]
  15. Ricquier D., Lin C., Klingenberg M. Isolation of the GDP binding protein from brown adipose tissue mitochondria of several animals and amino acid composition study in rat. Biochem Biophys Res Commun. 1982 May 31;106(2):582–589. doi: 10.1016/0006-291x(82)91150-0. [DOI] [PubMed] [Google Scholar]
  16. Ricquier D., Mory G., Hemon P. Changes induced by cold adaptation in the brown adipose tissue from several species of rodents, with special reference to the mitochondrial components. Can J Biochem. 1979 Nov;57(11):1262–1266. doi: 10.1139/o79-167. [DOI] [PubMed] [Google Scholar]
  17. Ricquier D., Nechad M., Mory G. Ultrastructural and biochemical characterization of human brown adipose tissue in pheochromocytoma. J Clin Endocrinol Metab. 1982 Apr;54(4):803–807. doi: 10.1210/jcem-54-4-803. [DOI] [PubMed] [Google Scholar]
  18. Sebald W., Wild G. Mitochondrial ATPase complex from Neurospora crassa. Methods Enzymol. 1979;55:344–351. doi: 10.1016/0076-6879(79)55043-5. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES