Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1988 Jan 15;249(2):451–457. doi: 10.1042/bj2490451

Regulation of GDP binding and uncoupling-protein concentration in brown-adipose-tissue mitochondria. The effects of cold-acclimation, warm-reacclimation and noradrenaline.

T Peachey 1, R R French 1, D A York 1
PMCID: PMC1148724  PMID: 3342023

Abstract

We have used a specific immunoassay for uncoupling protein and [3H]GDP binding to study the acute and chronic responses of brown-adipose-tissue (BAT) mitochondria of warm-acclimated rats to housing at 4 degrees C and cold-acclimated rats to housing at 27 degrees C. These studies have shown the following. (1) In the cold-exposed rat the increase in mitochondrial uncoupling-protein concentration parallels the increase in GDP binding from 1 day to 5 days, but that acutely (initial 4 h) the increase in GDP binding is not associated with any change in uncoupling-protein concentration. 2. In the cold-acclimated rat rehoused at 27 degrees C, GDP binding fell by over 50% in the first 2 days, without any change in uncoupling-protein concentrations. 3. Noradrenaline acutely (30 min) increased BAT mitochondrial GDP binding of lean and obese Zucker rats, without any change in uncoupling-protein concentrations. 4. The increases in GDP binding in cold-exposed rats were associated with increases in the rate of swelling of mitochondria in the presence of valinomycin and potassium acetate. The evidence supports the hypothesis that the acute response of the rat to changes in environmental temperature are associated with unmasking or remasking of uncoupling protein, whereas chronically changes in uncoupling-protein concentration predominate.

Full text

PDF
453

Selected References

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

  1. Ashwell M., Holt S., Jennings G., Stirling D. M., Trayhurn P., York D. A. Measurement by radioimmunoassay of the mitochondrial uncoupling protein from brown adipose tissue of obese (ob/ob) mice and Zucker (fa/fa) rats at different ages. FEBS Lett. 1985 Jan 7;179(2):233–237. doi: 10.1016/0014-5793(85)80525-1. [DOI] [PubMed] [Google Scholar]
  2. Ashwell M., Jennings G., Richard D., Stirling D. M., Trayhurn P. Effect of acclimation temperature on the concentration of the mitochondrial 'uncoupling' protein measured by radioimmunoassay in mouse brown adipose tissue. FEBS Lett. 1983 Sep 5;161(1):108–112. doi: 10.1016/0014-5793(83)80740-6. [DOI] [PubMed] [Google Scholar]
  3. Cannon B., Lindberg O. Mitochondria from brown adipose tissue: isolation and properties. Methods Enzymol. 1979;55:65–78. doi: 10.1016/0076-6879(79)55010-1. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Desautels M. Mitochondrial thermogenin content is unchanged during atrophy of BAT of fasting mice. Am J Physiol. 1985 Jul;249(1 Pt 1):E99–106. doi: 10.1152/ajpendo.1985.249.1.E99. [DOI] [PubMed] [Google Scholar]
  6. Desautels M., Zaror-Behrens G., Himms-Hagen J. Increased purine nucleotide binding, altered polypeptide composition, and thermogenesis in brown adipose tissue mitochondria of cold-acclimated rats. Can J Biochem. 1978 Jun;56(6):378–383. doi: 10.1139/o78-060. [DOI] [PubMed] [Google Scholar]
  7. Foster D. O., Frydman M. L. Nonshivering thermogenesis in the rat. II. Measurements of blood flow with microspheres point to brown adipose tissue as the dominant site of the calorigenesis induced by noradrenaline. Can J Physiol Pharmacol. 1978 Feb;56(1):110–122. doi: 10.1139/y78-015. [DOI] [PubMed] [Google Scholar]
  8. French R. R., Holt S. J., York D. A. High affinity GDP binding sites on brown adipose tissue mitochondria of genetically obese 'fa/fa' rats. Biosci Rep. 1985 Feb;5(2):159–166. doi: 10.1007/BF01117062. [DOI] [PubMed] [Google Scholar]
  9. Gribskov C. L., Henningfield M. F., Swick A. G., Swick R. W. Evidence for unmasking of rat brown-adipose-tissue mitochondrial GDP-binding sites in response to acute cold exposure. Effects of washing with albumin on GDP binding. Biochem J. 1986 Feb 1;233(3):743–747. doi: 10.1042/bj2330743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Heaton G. M., Wagenvoord R. J., Kemp A., Jr, Nicholls D. G. Brown-adipose-tissue mitochondria: photoaffinity labelling of the regulatory site of energy dissipation. Eur J Biochem. 1978 Jan 16;82(2):515–521. doi: 10.1111/j.1432-1033.1978.tb12045.x. [DOI] [PubMed] [Google Scholar]
  11. Hogan S., Himms-Hagen J. Abnormal brown adipose tissue in obese (ob/ob) mice: response to acclimation to cold. Am J Physiol. 1980 Oct;239(4):E301–E309. doi: 10.1152/ajpendo.1980.239.4.E301. [DOI] [PubMed] [Google Scholar]
  12. Holloway P. W. A simple procedure for removal of Triton X-100 from protein samples. Anal Biochem. 1973 May;53(1):304–308. doi: 10.1016/0003-2697(73)90436-3. [DOI] [PubMed] [Google Scholar]
  13. Klingenberg M., Winkler E. The reconstituted isolated uncoupling protein is a membrane potential driven H+ translocator. EMBO J. 1985 Dec 1;4(12):3087–3092. doi: 10.1002/j.1460-2075.1985.tb04049.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Lean M. E., Branch W. J., James W. P., Jennings G., Ashwell M. Measurement of rat brown-adipose-tissue mitochondrial uncoupling protein by radioimmunoassay: increased concentration after cold acclimation. Biosci Rep. 1983 Jan;3(1):61–71. doi: 10.1007/BF01121572. [DOI] [PubMed] [Google Scholar]
  17. Lin C. S., Hackenberg H., Klingenberg E. M. The uncoupling protein from brown adipose tissue mitochondria is a dimer. A hydrodynamic study. FEBS Lett. 1980 May 5;113(2):304–306. doi: 10.1016/0014-5793(80)80614-4. [DOI] [PubMed] [Google Scholar]
  18. Lin C. S., Klingenberg M. Characteristics of the isolated purine nucleotide binding protein from brown fat mitochondria. Biochemistry. 1982 Jun 8;21(12):2950–2956. doi: 10.1021/bi00541a023. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Mory G., Bouillaud F., Combes-George M., Ricquier D. Noradrenaline controls the concentration of the uncoupling protein in brown adipose tissue. FEBS Lett. 1984 Jan 30;166(2):393–396. doi: 10.1016/0014-5793(84)80120-9. [DOI] [PubMed] [Google Scholar]
  21. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  22. Nedergaard J., Cannon B. Apparent unmasking of [3H]GDP binding in rat brown-fat mitochondria is due to mitochondrial swelling. Eur J Biochem. 1987 May 4;164(3):681–686. doi: 10.1111/j.1432-1033.1987.tb11180.x. [DOI] [PubMed] [Google Scholar]
  23. Nedergaard J., Cannon B. [3H]GDP binding and thermogenin amount in brown adipose tissue mitochondria from cold-exposed rats. Am J Physiol. 1985 Mar;248(3 Pt 1):C365–C371. doi: 10.1152/ajpcell.1985.248.3.C365. [DOI] [PubMed] [Google Scholar]
  24. Nedergaard J., Raasmaja A., Cannon B. Parallel increases in amount of (3H)GDP binding and thermogenin antigen in brown-adipose-tissue mitochondria of cafeteria-fed rats. Biochem Biophys Res Commun. 1984 Aug 16;122(3):1328–1336. doi: 10.1016/0006-291x(84)91237-3. [DOI] [PubMed] [Google Scholar]
  25. Nicholls D. G. Hamster brown-adipose-tissue mitochondria. Purine nucleotide control of the ion conductance of the inner membrane, the nature of the nucleotide binding site. Eur J Biochem. 1976 Feb 16;62(2):223–228. doi: 10.1111/j.1432-1033.1976.tb10151.x. [DOI] [PubMed] [Google Scholar]
  26. Nicholls D. G., Lindberg O. Brown-adipose-tissue mitochondria. The influence of albumin and nucleotides on passive ion permeabilities. Eur J Biochem. 1973 Sep 3;37(3):523–530. doi: 10.1111/j.1432-1033.1973.tb03014.x. [DOI] [PubMed] [Google Scholar]
  27. Nicholls D. G., Locke R. M. Thermogenic mechanisms in brown fat. Physiol Rev. 1984 Jan;64(1):1–64. doi: 10.1152/physrev.1984.64.1.1. [DOI] [PubMed] [Google Scholar]
  28. Rial E., Nicholls D. G. The mitochondrial uncoupling protein from guinea-pig brown adipose tissue. Synchronous increase in structural and functional parameters during cold-adaptation. Biochem J. 1984 Sep 15;222(3):685–693. doi: 10.1042/bj2220685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ricquier D., Bouillaud F., Toumelin P., Mory G., Bazin R., Arch J., Pénicaud L. Expression of uncoupling protein mRNA in thermogenic or weakly thermogenic brown adipose tissue. Evidence for a rapid beta-adrenoreceptor-mediated and transcriptionally regulated step during activation of thermogenesis. J Biol Chem. 1986 Oct 25;261(30):13905–13910. [PubMed] [Google Scholar]
  30. 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]
  31. Swick A. G., Swick R. W. Rapid changes in number of GDP binding sites on brown adipose tissue mitochondria. Am J Physiol. 1986 Aug;251(2 Pt 1):E192–E195. doi: 10.1152/ajpendo.1986.251.2.E192. [DOI] [PubMed] [Google Scholar]
  32. Trayhurn P., Jennings G. Evidence that fasting can induce a selective loss of uncoupling protein from brown adipose tissue mitochondria of mice. Biosci Rep. 1986 Sep;6(9):805–810. doi: 10.1007/BF01117103. [DOI] [PubMed] [Google Scholar]
  33. Trayhurn P., Richard D., Jennings G., Ashwell M. Adaptive changes in the concentration of the mitochondrial 'uncoupling' protein in brown adipose tissue of hamsters acclimated at different temperatures. Biosci Rep. 1983 Dec;3(12):1077–1084. doi: 10.1007/BF01120199. [DOI] [PubMed] [Google Scholar]
  34. York D. A., Marchington D., Holt S. J., Allars J. Regulation of sympathetic activity in lean and obese Zucker (fa/fa) rats. Am J Physiol. 1985 Sep;249(3 Pt 1):E299–E305. doi: 10.1152/ajpendo.1985.249.3.E299. [DOI] [PubMed] [Google Scholar]
  35. Young J. B., Saville E., Rothwell N. J., Stock M. J., Landsberg L. Effect of diet and cold exposure on norepinephrine turnover in brown adipose tissue of the rat. J Clin Invest. 1982 May;69(5):1061–1071. doi: 10.1172/JCI110541. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES