[3H]GDP binding and thermogenin amount in brown adipose tissue mitochondria from cold-exposed rats
- 1 March 1985
- journal article
- Published by American Physiological Society in American Journal of Physiology-Cell Physiology
- Vol. 248 (3), C365-C371
- https://doi.org/10.1152/ajpcell.1985.248.3.c365
Abstract
Brown fat mitochondria were isolated from cold-exposed and control rats, and their content of the brown-fat-specific 32-kDa “uncoupling” protein thermogenin determined both by the traditional [3H]GDP-binding method and by the recently developed enzyme-linked immunosorbent assay (ELISA). In mitochondria isolated from both cold-acclimated (3 wk at 4 degrees C) and cold-exposed rats (24 h), an increase in thermogenin content was observable, both when estimated by the [3H]GDP-binding method and by the ELISA assay, and there was no statistically significant difference in the magnitude of these increases in the two methods. In 1 h cold-exposed rats there was no increase in [3H]GDP binding or in the ELISA reaction. When the amount of thermogenin was plotted against [3H]GDP binding in the different states, a relationship of 75,000 g thermogenin per mole GDP bound was obtained. Based on the resolution of these two methods, and under the three conditions investigated, it was concluded that there was no reason to postulate the existence of a “masked” form of thermogenin or of an “unmasking” process and that thermogenin in the mitochondria, as in the isolated state, has apparently one GDP binding site per dimer.Keywords
This publication has 11 references indexed in Scilit:
- Effects of Dietary Essential Fatty Acids on Active Thermogenin Content in Rat Brown Adipose TissueJournal of Nutrition, 1983
- An immunological study of the uncoupling protein of brown adipose tissue mitochondriaBiochemical Journal, 1983
- Biochemical aspects of acclimation to coldJournal of Thermal Biology, 1983
- Measurement of rat brown-adipose-tissue mitochondrial uncoupling protein by radioimmunoassay: Increased concentration after cold acclimationBioscience Reports, 1983
- Characteristics of the isolated purine nucleotide binding protein from brown fat mitochondriaBiochemistry, 1982
- The Brown Fat CellInternational Review of Cytology, 1982
- Roles of noradrenaline and protein synthesis in the cold-induced increase in purine nucleotide binding by rat brown adipose tissue mitochondriaCanadian Journal of Biochemistry, 1979
- [8] Mitochondria from brown adipose tissue: Isolation and propertiesMethods in Enzymology, 1979
- Increased purine nucleotide binding, altered polypeptide composition, and thermogenesis in brown adipose tissue mitochondria of cold-acclimated ratsCanadian Journal of Biochemistry, 1978
- Mitochondrial protein alteration in active brown fat: A sodium dodecyl sulfate-polyacrylamide gel electrophoretic studyBiochemical and Biophysical Research Communications, 1976