Advertisement

Molecular and Cellular Biochemistry

, Volume 92, Issue 2, pp 147–157 | Cite as

Microsomal redox systems in brown adipose tissue: high lipid peroxidation, low cholesterol biosynthesis and no detectable cytochrome P-450

  • B. Seshadri Sekhar
  • C. K. Ramakrishna Kurup
  • T. Ramasarma
Original Article

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 b5 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.

Key words

microsomal redox systems brown adipose tissue lipid peroxidation cholesterol biosynthesis cytochrome P-450 

Abbreviations

BAT

Brown Adipose Tissue

MDA

malondialdehyde

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Smith RE, Horwitz BA: Brown fat and thermogenesis. Physiol Rev 49: 330–425, 1969Google Scholar
  2. 2.
    Flatmark T, Pederson JI: Brown adipose tissue mitochondria. Biochim Biophys Acta 416: 53–103, 1975Google Scholar
  3. 3.
    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, 1978Google Scholar
  4. 4.
    Lindberg O, Cannon B, Nedergaard J: Thermogenic mitochondria, in ‘Mitochondria and Microsomes’ (Lee CP et al., eds). Addison-Wiley, Reading, MA, USA 1981, pp 93–119Google Scholar
  5. 5.
    Cannon B, Nedergaard J: The biochemistry of an inefficient tissue: Brown Adipose Tissue. Essays Biochem 20: 110–163, 1985Google Scholar
  6. 6.
    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, 1979Google Scholar
  7. 7.
    Lin C, Klingenberg M: Characteristics of the isolated purine nucleotide binding protein from brown fat mitochondria. Biochemistry 21: 2950–2956, 1982Google Scholar
  8. 8.
    Nicholls D: Brown Adipose Tissue Mitochondria. Biochim Biophys Acta 549: 1–29, 1979Google Scholar
  9. 9.
    Cannon B, Lindberg O: Mitochondria from Brown Adipose Tissue: Isolation and properties. Methods Enzymol 5517: 65–78, 1979Google Scholar
  10. 10.
    Seshadri Sekhar B, Ramakrishna Kurup CK, Ramasarma T: Generation of hydrogen peroxide by Brown Adipose Tissue Mitochondria. J Bioenergetics Biomemb 19: 397–407, 1987Google Scholar
  11. 11.
    Hochstein P, Nordebrand K, Ernster L: ADP-activated lipid peroxidation coupled to TPNH oxidase system of microsomes. Biochem Biophys Res Commun 14: 323–328, 1964Google Scholar
  12. 12.
    Wills ED: Lipid peroxidation formation in microsomesGeneral considerations. Biochem J 113: 325–332, 1969Google Scholar
  13. 13.
    Kappus H, Sies H: Toxic effects associated with oxygen metabolism: Redox cycling and lipid peroxidation. Experientia 37: 1233–1241, 1981Google Scholar
  14. 14.
    de Duve D: Evolution of the peroxisome. Ann NY Acad Sci 168: 369–381, 1969Google Scholar
  15. 15.
    Howirtz BA: Can J Physiol Pharmacol A54: 45–48, 1976Google Scholar
  16. 16.
    Ramasarma T: Generation of H2O2 in biomembranes. Biochim Biophys Acta 694: 69–93, 1982Google Scholar
  17. 17.
    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, 1984Google Scholar
  18. 18.
    Strittmatter P: NADH-cytochrome b5 reductase. Methods Enzymol 10: 561–565, 1967Google Scholar
  19. 19.
    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, 1967Google Scholar
  20. 20.
    Imai Y, Ito A, Sato R: Evidence for biochemically different types of vesicles in the hepatic microsomal fraction. J Biochem (Tokyo) 60: 417–428, 1966Google Scholar
  21. 21.
    Guengerich FP: In Principles and Methods of Toxicology (Hayes WA, ed). Raven Press, New York, USA, 1982, pp 609–634Google Scholar
  22. 22.
    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, 1963Google Scholar
  23. 23.
    Avigan J, Goodman DS, Steinberg D: Studies of cholesterol Biosynthesi. J Biol Chem 238: 1283–1286, 1963Google Scholar
  24. 24.
    Stadtman TC: Preparation and assay of cholesterol and ergosterol. Methods Enzymol 3: 392–394, 1957Google Scholar
  25. 25.
    Lester RL, Crane FL: The natural occurrence of coenzyme Q and related compound. J Biol Chem 234: 2169–2174, 1959Google Scholar
  26. 26.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurements with folin phenol reagent. J Biol Chem 193: 265–275, 1951PubMedGoogle Scholar
  27. 27.
    Estabrook RW, Werringloer J: Estimation of cytochrome P-450. Methods Enzymol 52: 212–220, 1978Google Scholar
  28. 28.
    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, 1976Google Scholar
  29. 29.
    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, 1976Google Scholar
  30. 30.
    Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 227: 680–685, 1970Google Scholar
  31. 31.
    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, 1984Google Scholar
  32. 32.
    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, 1967Google Scholar
  33. 33.
    Moore RW, Welton AF, Aust SD: Methods Enzymol 52: 324–331, 1978Google Scholar
  34. 34.
    Joel CD, Ball EG: The electron transmitter system of Brown Adipose Tissue. Biochemistry 1: 281–284, 1962Google Scholar
  35. 35.
    Sivaramakrishnan S, Ramasarma T: Activation of succinate dehydrogenase in Brown Adipose Tissue Mitochondria. Indian J Biochem Biophys 15: 14–18, 1978Google Scholar
  36. 36.
    Ernster L, Nordenbrand K: Microsomal lipid peroxidation. Methods Enzymol 10: 574–580, 1967Google Scholar
  37. 37.
    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, 1964Google Scholar
  38. 38.
    Kappus H: Lipid peroxidation; Mechanisms, Analysis, Enzymology and Biological Relevance in ‘Oxidative stress’ (Sies H ed) Academic Press, London, 1985, pp 273–310Google Scholar
  39. 39.
    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, 1973Google Scholar
  40. 40.
    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 225–233Google Scholar

Copyright information

© Kluwer Academic Publishers 1990

Authors and Affiliations

  • B. Seshadri Sekhar
    • 1
  • C. K. Ramakrishna Kurup
    • 1
  • T. Ramasarma
    • 1
  1. 1.The Department of BiochemistryIndian Institute of ScienceBangaloreIndia

Personalised recommendations