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Δ5 Desaturase activity in rat kidney microsomes

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Abstract

Rat kidney microsomal fraction is able to catalyze the enzymatic desaturation of eicosatrienoic acid (20:3n-6) to arachidonic acid (20:4n-6) by the Δ5 desaturase pathway, in the presence of reduced nicotinamide adenine dinucleotide (NADH), adenosinetriphosphate (ATP) and coenzyme A (CoA). The substrate of the reaction [1-14C]eicosa-8,11,14trienoic acid (20:3n-6), was separated from the product [1-14C]eicosa-5,8,11,14-tetraenoic acid (20:4n-6) by reverse phase high-pressure liquid chromatography (RP-HPLC). These fatty acids were individually collected by monitoring the eluent at 205 nm and their radioactivity was measured by liquid scintillation counting. The Δ5 desaturase activity in kidney microsomes increased linearly with the substrate concentration up to 20 μM. Enzymatic activity was sensitive to pH with the maximum at 7.0 and was proportional with incubation time up to 10 min. The apparent Km and Vmax of Δ5 desaturase were 56 μM and 60 pmoles·min−1·mg−1 microsomal protein, respectively. Neither the cytosolic renal fraction nor the cytosolic liver fraction enhanced the Δ5 desaturase activity. Contrary to a report but in accordance to others, the present results suggest that rat kidneys can synthesize arachidonic acid at least to satisfy partially their needs for eicosanoid production.

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References

  1. Strittmatter P, Spatz L, Corcoran D, Rogers MJ, Setlow B, Redline R: Purification and properties of rat liver microsomal stearyl-CoA desaturase. Proc Natl Acad Sci USA 71: 4565–4569, 1974

    Google Scholar 

  2. Okayasu T, Nagao M, Ishibashi T, Imai Y: Purification and partial characterization of linoleoyl-CoA desaturase from rat liver microsomes. Archs Biochem Biophys 206: 21–28, 1981

    Google Scholar 

  3. Holloway PW: A requirement for three protein components in microsomal stearyl coenzyme A desaturation. Biochemistry 10: 1556–1560, 1971

    Google Scholar 

  4. Brenner RR: Regulatory functions of Δ6 desaturase. Key enzyme of polyunsaturated fatty acid biosynthesis. In: NG Bazán, RR Brenner, NM Giusto (eds) Function and Biosynthesis of Lipids. Adv Exp Med Biol Vol 83 Plenum Press, New York and London, 1977, pp 85–101

    Google Scholar 

  5. Brenner RR: Factors influencing fatty acid chain elongation and desaturation. In: AG Vergroesen, M Crawford (eds) The Role of Fast in Human Nutrition Academic Press, London, 1989, pp 45–79.

    Google Scholar 

  6. Cinti DL, Montgomery MR: Stimulation of hepatic cytochrome b5-mediated lipid desaturation by renal microsomes. Life Sci 18: 1223–1228, 1976

    Google Scholar 

  7. Suneja SK, Nagi MN, Cook L, Osei P, Cinti DL: Do rat kidney cortex microsomes possess the enzymatic machinery to desaturate and chain elongate fatty acyl-CoA derivatives? Lipids 26: 359–363, 1991

    Google Scholar 

  8. Mandon EC, Gómez Dumm INT de, Brenner RR Effect of high carbohydrate and high protein diets on microsomal fatty acid composition, ‘fluidity’ and Δ6 desaturation activity in kidney and lung. Acta Physiol Pharmacol Latinoam 38: 49–58, 1988

    Google Scholar 

  9. Chern JC, Kinsella JE: Concurrent release and reacylation of fatty acids by rat kidney cells. Selective incorporation of arachidonic acid. Biochim Biophys Acta 750: 7–17, 1983

    Google Scholar 

  10. McGiff JC: New developments in prostaglandin and thromboxane research. Fed Proc 38: 64–65, 1979

    Google Scholar 

  11. Willis AL: Nutritional and pharmacological factors in eicosanoid biology. Nutr Rev 39: 289–295, 1981

    Google Scholar 

  12. Schwartzman ML, Abraham NG, Carroll MA, Levere RD, McGiff JC: Regulation of arachidonic acid metabolism by cytochrome P-450 in rabbit kidney. Biochem J 238: 283–290, 1986

    Google Scholar 

  13. Lefkowith JB, Flippo V, Sprecher H, Needleman P: Paradoxical conservation of cardiac and renal arachidonate content in essential fatty acid deficiency. J Biol Chem 260: 15736–15744, 1985

    Google Scholar 

  14. Montgomery MR, Cinti DL: Pyridine nucleotide-dependent electron transport in kidney cortex microsomes: Interaction with desaturase and other microsomal mixed-function oxidases. Mol Pharmacol 13: 60–69, 1977

    Google Scholar 

  15. Cinti DL, Cook L, Nagi MN, Sanoj KS: The fatty acid chain elongation system ofmmammalian endoplasmic reticulum. Prog Lipid Res 31: 1–51, 1992

    Google Scholar 

  16. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  17. Garda HA, Leikin AI, Brenner RR: Determination of fatty acid desaturation by RP-HPLC. Anal Asoc Quím Arg 80: 365–371, 1992

    Google Scholar 

  18. González-Baró M, Irazú CE, Pollero RJ: Palmitoyl-CoA ligase activity in hepatopancreas and gill microsomes of the freshwater shrimpMacrobrachium borellii. Comp Biochem Physiol 97B: 129–133, 1990

    Google Scholar 

  19. Narce M, Gresti J, Bézard J: Methods for evaluating the bioconversion of radioactive polyunsaturated fatty acids by use of reversed-phase liquid chromatography. J Chromatog 448: 249–264, 1988

    Google Scholar 

  20. Kawata S, Chitranukroh A, Owen JS, McIntyre N: Membrane lipid changes in erythrocytes, liver and kidney in acute and chronic experimental liver disease in rats. Biochim Biophys Acta 896: 26–34, 1987

    Google Scholar 

  21. Brenner RR: The desaturation step in the animal biosynthesis of polyunsaturated fatty acids. Lipids 6: 567–575, 1971

    Google Scholar 

  22. Brenner RR, Peluffo RO: Effect of saturated and unsaturated fatty acids on the desaturation ‘in vitro’ of palmitic, stearic, oleic, linoleic and linolenic acid. J Biol Chem 241: 5213–5219, 1966

    Google Scholar 

  23. Nervi AM, Brenner RR, Peluffo RO: Effect of arachidonic acid on the microsomal desaturation of linoleic into γ-linolenic acid and their simultaneous incorporation into the phospholipids. Biochim Biophys Acta 152: 539–551, 1968

    Google Scholar 

  24. Blond JP, Bézard J: Δ5 Desaturation of dihomogamma linolenic acid (20:3(n-6)) into arachidonic acid (20:4(n-6)) by rat liver microsomes and incorporation of fatty acids in microsome phospholipids. Biochim Biophys Acta 1084: 255–260, 1991

    Google Scholar 

  25. Yamoaka K, Okayasu T, Ishibashi T: Product identification and determination of optimal condition of 8,11,14-eicosatrienoic acid desaturation in rat liver microsomes. Hokkaido Igaku Zasshi 61: 755–765, 1986

    Google Scholar 

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Authors and Affiliations

  1. Facultad de Ciencias Médicas, Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP), 60 y 120, 1900, La Plata, Argentina

    Carlos E. Irazú, Susana González-Rodríguez & Rodolfo R. Brenner

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  1. Carlos E. Irazú
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  2. Susana González-Rodríguez
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  3. Rodolfo R. Brenner
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Irazú, C.E., González-Rodríguez, S. & Brenner, R.R. Δ5 Desaturase activity in rat kidney microsomes. Mol Cell Biochem 129, 31–37 (1993). https://doi.org/10.1007/BF00926573

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  • DOI: https://doi.org/10.1007/BF00926573

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