Publications

• Elongase reactions as control points in long-chain polyunsaturated fatty acid synthesis.

  Gregory M.K., Gibson R.A., Cook-Johnson R.J., Cleland L.G., James M.J.

  <h4>Background</h4>Δ6-Desaturase (Fads2) is widely regarded as rate-limiting in the conversion of dietary α-linolenic acid (18:3n-3; ALA) to the long-chain omega-3 polyunsaturated fatty acid docosahexaenoic acid (22:6n-3; DHA). However, increasing dietary ALA or the direct Fads2 product, stearidon ... >> More

  <h4>Background</h4>Δ6-Desaturase (Fads2) is widely regarded as rate-limiting in the conversion of dietary α-linolenic acid (18:3n-3; ALA) to the long-chain omega-3 polyunsaturated fatty acid docosahexaenoic acid (22:6n-3; DHA). However, increasing dietary ALA or the direct Fads2 product, stearidonic acid (18:4n-3; SDA), increases tissue levels of eicosapentaenoic acid (20:5n-3; EPA) and docosapentaenoic acid (22:5n-3; DPA), but not DHA. These observations suggest that one or more control points must exist beyond ALA metabolism by Fads2. One possible control point is a second reaction involving Fads2 itself, since this enzyme catalyses desaturation of 24:5n-3 to 24:6n-3, as well as ALA to SDA. However, metabolism of EPA and DPA both require elongation reactions. This study examined the activities of two elongase enzymes as well as the second reaction of Fads2 in order to concentrate on the metabolism of EPA to DHA.<h4>Methodology/principal findings</h4>The substrate selectivities, competitive substrate interactions and dose response curves of the rat elongases, Elovl2 and Elovl5 were determined after expression of the enzymes in yeast. The competitive substrate interactions for rat Fads2 were also examined. Rat Elovl2 was active with C(20) and C(22) polyunsaturated fatty acids and this single enzyme catalysed the sequential elongation reactions of EPA→DPA→24:5n-3. The second reaction DPA→24:5n-3 appeared to be saturated at substrate concentrations not saturating for the first reaction EPA→DPA. ALA dose-dependently inhibited Fads2 conversion of 24:5n-3 to 24:6n-3.<h4>Conclusions</h4>The competition between ALA and 24:5n-3 for Fads2 may explain the decrease in DHA levels observed after certain intakes of dietary ALA have been exceeded. In addition, the apparent saturation of the second Elovl2 reaction, DPA→24:5n-3, provides further explanations for the accumulation of DPA when ALA, SDA or EPA is provided in the diet. This study suggests that Elovl2 will be critical in understanding if DHA synthesis can be increased by dietary means. << Less

  PLoS ONE 6:E29662-E29662(2011) [PubMed] [EuropePMC]

  This publication is cited by 6 other entries.

• The same rat Delta6-desaturase not only acts on 18- but also on 24-carbon fatty acids in very-long-chain polyunsaturated fatty acid biosynthesis.

  D'andrea S., Guillou H., Jan S., Catheline D., Thibault J.N., Bouriel M., Rioux V., Legrand P.

  The recently cloned Delta6-desaturase is known to catalyse the first step in very-long-chain polyunsaturated fatty acid biosynthesis, i.e. the desaturation of linoleic and alpha-linolenic acids. The hypothesis that this enzyme could also catalyse the terminal desaturation step, i.e. the desaturati ... >> More

  The recently cloned Delta6-desaturase is known to catalyse the first step in very-long-chain polyunsaturated fatty acid biosynthesis, i.e. the desaturation of linoleic and alpha-linolenic acids. The hypothesis that this enzyme could also catalyse the terminal desaturation step, i.e. the desaturation of 24-carbon highly unsaturated fatty acids, has never been elucidated. To test this hypothesis, the activity of rat Delta6-desaturase expressed in COS-7 cells was investigated. Recombinant Delta6-desaturase expression was analysed by Western blot, revealing a single band at 45 kDa. The putative involvement of this enzyme in the Delta6-desaturation of C(24:5) n-3 to C(24:6) n-3 was measured by incubating transfected cells with C(22:5) n-3. Whereas both transfected and non-transfected COS-7 cells were able to synthesize C(24:5) n-3 by elongation of C(22:5) n-3, only cells expressing Delta6-desaturase were also able to produce C(24:6) n-3. In addition, Delta6-desaturation of [1-(14)C]C(24:5) n-3 was assayed in vitro in homogenates from COS-7 cells expressing Delta6-desaturase or not, showing that Delta6-desaturase catalyses the conversion of C(24:5) n-3 to C(24:6) n-3. Evidence is therefore presented that the same rat Delta6-desaturase catalyses not only the conversion of C(18:3) n-3 to C(18:4) n-3, but also the conversion of C(24:5) n-3 to C(24:6) n-3. A similar mechanism in the n-6 series is strongly suggested. << Less

  Biochem. J. 364:49-55(2002) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Trans-vaccenate is Delta13-desaturated by FADS3 in rodents.

  Rioux V., Pedrono F., Blanchard H., Duby C., Boulier-Monthean N., Bernard L., Beauchamp E., Catheline D., Legrand P.

  Fatty acid desaturases play critical roles in regulating the biosynthesis of unsaturated fatty acids in all biological kingdoms. As opposed to plants, mammals are so far characterized by the absence of desaturases introducing additional double bonds at the methyl-end site of fatty acids. However, ... >> More

  Fatty acid desaturases play critical roles in regulating the biosynthesis of unsaturated fatty acids in all biological kingdoms. As opposed to plants, mammals are so far characterized by the absence of desaturases introducing additional double bonds at the methyl-end site of fatty acids. However, the function of the mammalian fatty acid desaturase 3 (FADS3) gene remains unknown. This gene is located within the FADS cluster and presents a high nucleotide sequence homology with FADS1 (Δ5-desaturase) and FADS2 (Δ6-desaturase). Here, we show that rat FADS3 displays no common Δ5-, Δ6- or Δ9-desaturase activity but is able to catalyze the unexpected Δ13-desaturation of trans-vaccenate. Although there is no standard for complete conclusive identification, structural characterization strongly suggests that the Δ11,13-conjugated linoleic acid (CLA) produced by FADS3 from trans-vaccenate is the trans11,cis13-CLA isomer. In rat hepatocytes, knockdown of FADS3 expression specifically reduces trans-vaccenate Δ13-desaturation. Evidence is presented that FADS3 is the first "methyl-end" fatty acid desaturase functionally characterized in mammals. << Less

  J. Lipid Res. 54:3438-3452(2013) [PubMed] [EuropePMC]

  This publication is cited by 4 other entries.