Abstract
The presence of redox systems in microsomes of brown adipose tissue (BAT) in cold exposed rats was investigated and compared with liver. BAT microsomes showed high activity of lipid peroxidation measured both by the formation of malondialdehyde (MDA) and by oxygen uptake. NADH and NADPH dependent cytochrome c reductase activity were present in both BAT and liver microsomes. Aminopyrine demethylase and aniline hydroxylase activities, the characteristic detoxification enzymes in liver microsomes could not be detected in BAT microsomes. BAT minces showed very poor incorporation of [1-14C]acetate and [2-14C]-mevalonate in unsaponifiable lipid fraction compared to liver. Biosynthesis of cholesterol and ubiquinone, but not fatty acids, and the activity of 3-hydroxy-3-methyl glutaryl CoA reductase appear to be very low in BAT. Examination of difference spectra showed the presence of only cytochrome b 5 in BAT microsomes. In addition to the inability to detect the enzyme activities dependent on cytochrome P-450, a protein with the characteristic spectrum, molecular size in SDS-PAGE and interaction with antibodies in double diffusion test, also could not be detected in BAT microsomes. The high activity of lipid peroxidation in microsomes, being associated with large oxygen uptake and oxidation of NADPH, will also contribute to the energy dissipation as heat in BAT, considered important in thermogenesis.
Similar content being viewed by others
Abbreviations
- BAT:
-
Brown Adipose Tissue
- MDA:
-
malondialdehyde
References
Smith RE, Horwitz BA: Brown fat and thermogenesis. Physiol Rev 49: 330–425, 1969
Flatmark T, Pederson JI: Brown adipose tissue mitochondria. Biochim Biophys Acta 416: 53–103, 1975
Foster DO, Frydman ML: Tissue distribution of cold-induced thermogenesis in conscious warm- and cold-acclimated rats reevaluated from changes in tissue blood flow. The dominant role of brown adipose tissue in the replacement of shivering by nonshivering thermogenesis. Can J Physiol Pharmacol 56: 110–122, 1978
Lindberg O, Cannon B, Nedergaard J: Thermogenic mitochondria, in ‘Mitochondria and Microsomes’ (Lee CP et al., eds). Addison-Wiley, Reading, MA, USA 1981, pp 93–119
Cannon B, Nedergaard J: The biochemistry of an inefficient tissue: Brown Adipose Tissue. Essays Biochem 20: 110–163, 1985
Ricqier D, Gerrais C, Kader JC, Hemon P: Partial purification by Guanosine-5′-diphosphate-agarose affinity chromatography of the 32000 molecular wt polypeptide from mitochondria of Brown Adipose Tissue. FEBS Lett 101: 35–38, 1979
Lin C, Klingenberg M: Characteristics of the isolated purine nucleotide binding protein from brown fat mitochondria. Biochemistry 21: 2950–2956, 1982
Nicholls D: Brown Adipose Tissue Mitochondria. Biochim Biophys Acta 549: 1–29, 1979
Cannon B, Lindberg O: Mitochondria from Brown Adipose Tissue: Isolation and properties. Methods Enzymol 5517: 65–78, 1979
Seshadri Sekhar B, Ramakrishna Kurup CK, Ramasarma T: Generation of hydrogen peroxide by Brown Adipose Tissue Mitochondria. J Bioenergetics Biomemb 19: 397–407, 1987
Hochstein P, Nordebrand K, Ernster L: ADP-activated lipid peroxidation coupled to TPNH oxidase system of microsomes. Biochem Biophys Res Commun 14: 323–328, 1964
Wills ED: Lipid peroxidation formation in microsomesGeneral considerations. Biochem J 113: 325–332, 1969
Kappus H, Sies H: Toxic effects associated with oxygen metabolism: Redox cycling and lipid peroxidation. Experientia 37: 1233–1241, 1981
de Duve D: Evolution of the peroxisome. Ann NY Acad Sci 168: 369–381, 1969
Howirtz BA: Can J Physiol Pharmacol A54: 45–48, 1976
Ramasarma T: Generation of H2O2 in biomembranes. Biochim Biophys Acta 694: 69–93, 1982
Ramasarma T, Maukassah Kelly S, Hochstein P: Inhibition of microsomal lipid peroxidation by cytosolic protein in presence of ADP and high concentration of Fe2+ Biochim Biophys Acta 796: 243–250, 1984
Strittmatter P: NADH-cytochrome b5 reductase. Methods Enzymol 10: 561–565, 1967
Masters BSS, Williams CH Jr, Kamin H: The preparation and properties of microsomal TPNH-cytochrome c reductase from pig liver. Methods Enzymol 10: 565–573, 1967
Imai Y, Ito A, Sato R: Evidence for biochemically different types of vesicles in the hepatic microsomal fraction. J Biochem (Tokyo) 60: 417–428, 1966
Guengerich FP: In Principles and Methods of Toxicology (Hayes WA, ed). Raven Press, New York, USA, 1982, pp 609–634
Joshi VC, Jayaraman J, Ramasarma T: Incorporation of mevalonic acid-2-C14 into ubichromenol and coenzyme Q in rat. Indian J Exptl Biol 1: 113–123, 1963
Avigan J, Goodman DS, Steinberg D: Studies of cholesterol Biosynthesi. J Biol Chem 238: 1283–1286, 1963
Stadtman TC: Preparation and assay of cholesterol and ergosterol. Methods Enzymol 3: 392–394, 1957
Lester RL, Crane FL: The natural occurrence of coenzyme Q and related compound. J Biol Chem 234: 2169–2174, 1959
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurements with folin phenol reagent. J Biol Chem 193: 265–275, 1951
Estabrook RW, Werringloer J: Estimation of cytochrome P-450. Methods Enzymol 52: 212–220, 1978
Matsubara T, Koike M, Touchi A, Yochimo A, Sugero L: Quantitative determination of cytochrome P-450 in rat liver homogenate. Anal Biochem 75: 596–603, 1976
Thomas PE, Lu AYH, Kawalek J, West SB, Levin W: Properties of electrophoretically homogeneous phenobarbital inducible and β-naphthoflavone-inducible forms of liver microsomal cytochrome P-450. J Biol Chem 251: 7929–7933, 1976
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 227: 680–685, 1970
Trazarkos JM, Bowen WD, Shafice A, Fischer RT, Gaylor JL: Cytochrome P-450 dependent oxidation of lanosterol in cholesterol biosynthesis. J Biol Chem 259: 13402–13412, 1984
Krishnaiah KV, Joshi VC, Ramasarma T: Effect of dietary cholesterol and coenzyme Q on the isoprene synthesis in rat liver. Arch Biochem Biophys 121: 147–153, 1967
Moore RW, Welton AF, Aust SD: Methods Enzymol 52: 324–331, 1978
Joel CD, Ball EG: The electron transmitter system of Brown Adipose Tissue. Biochemistry 1: 281–284, 1962
Sivaramakrishnan S, Ramasarma T: Activation of succinate dehydrogenase in Brown Adipose Tissue Mitochondria. Indian J Biochem Biophys 15: 14–18, 1978
Ernster L, Nordenbrand K: Microsomal lipid peroxidation. Methods Enzymol 10: 574–580, 1967
Orrenius S, Dallner G, Ernster L: Inhibition of the TPNH-linked lipid peroxidation of liver microsomes by drugs undergoing oxidative demethylation. Biochem Biophys Res Commun 14: 329–334, 1964
Kappus H: Lipid peroxidation; Mechanisms, Analysis, Enzymology and Biological Relevance in ‘Oxidative stress’ (Sies H ed) Academic Press, London, 1985, pp 273–310
Pederson TC, Buege JA, Aust SD: The role of reduced nicotinamide adenine dinucleotide phosphate-cytochrome c reductase in liver microsomal lipid peroxidation. J Biol Chem 248: 7134–7141, 1973
Ramasarma T, Seshadri Sekhar B, Ramakrishna Kurup CK: Cellular Thermogenesis: A New Approach in ‘Bio-oenergetics: Structure and Function of Energy Transducing Systems (Tozawa T, Papa S, eds) Japan Sci Soc Press Tokyo/Springer-Verlag, Berlin, 1987, pp 225–233
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Seshadri Sekhar, B., Ramakrishna Kurup, C.K. & Ramasarma, T. Microsomal redox systems in brown adipose tissue: high lipid peroxidation, low cholesterol biosynthesis and no detectable cytochrome P-450. Mol Cell Biochem 92, 147–157 (1990). https://doi.org/10.1007/BF00218132
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00218132