title: Function of human mitochondrial 2,4-dienoyl-CoA reductase and rat monofunctional Delta3-Delta2-enoyl-CoA isomerase in beta-oxidation of unsaturated fatty acids.
abstract: Human 2,4-dienoyl-CoA reductase (2,4-reductase; DECR) and rat monofunctional Delta(3)-Delta(2)-enoyl-CoA isomerase (rat 3, 2-isomerase; ECI) are thought to be mitochondrial auxiliary enzymes involved in the beta-oxidation of unsaturated fatty acids. However, their function during this process has not been demonstrated. Although they lack obvious peroxisomal targeting signals (PTSs), both proteins have been suggested previously to also occur in the mammalian peroxisomal compartment. The putative function and peroxisomal location of the two mammalian proteins can be examined in yeast, since beta-oxidation of unsaturated fatty acids is a compartmentalized process in Saccharomyces cerevisiae requiring peroxisomal 2,4-dienoyl-CoA reductase (Sps19p) and peroxisomal 3, 2-isomerase (Eci1p). A yeast sps19Delta mutant expressing human 2, 4-reductase ending with the native C-terminus could not grow on petroselinic acid [cis-C(18:1(6))] medium but could grow when the protein was extended with a PTS tripeptide, SKL (Ser-Lys-Leu). We therefore reason that the human protein is a physiological 2, 4-reductase but that it is probably not peroxisomal. Rat 3, 2-isomerase expressed in a yeast eci1Delta strain was able to re-establish growth on oleic acid [cis-C(18:1(9))] medium irrespective of an SKL extension. Since we had shown that Delta(2,4) double bonds could not be metabolized extra-peroxisomally to restore growth of the sps19Delta strain, we postulate that rat 3,2-isomerase acted on the Delta(3) unsaturated metabolite of oleic acid by replacing the mutant's missing activity from within the peroxisomes. Immunoblotting of fractionated yeast cells expressing rat 3, 2-isomerase in combination with electron microscopy supported our proposal that the protein functioned in peroxisomes. The results presented here shed new light on the function and location of human mitochondrial 2,4-reductase and rat monofunctional 3,2-isomerase.
authors.affiliation: Institut für Biochemie und Molekulare Zellbiologie der Universität Wien, Dr Bohrgasse 9, A-1030 Wien, Austria.
authors.forename: A
authors.initials: A
authors.lastname: Gurvitz
authors.forename: L
authors.initials: L
authors.lastname: Wabnegger
authors.forename: A I
authors.initials: AI
authors.lastname: Yagi
authors.forename: M
authors.initials: M
authors.lastname: Binder
authors.forename: A
authors.initials: A
authors.lastname: Hartig
authors.forename: H
authors.initials: H
authors.lastname: Ruis
authors.forename: B
authors.initials: B
authors.lastname: Hamilton
authors.forename: I W
authors.initials: IW
authors.lastname: Dawes
authors.forename: J K
authors.initials: JK
authors.lastname: Hiltunen
authors.forename: H
authors.initials: H
authors.lastname: Rottensteiner
mesh_terms: D000818:Animals; D019748:Carbon-Carbon Double Bond Isomerases / Q000378:metabolism*; D002455:Cell Division; D064006:Dodecenoyl-CoA Isomerase; D044943:Fatty Acid Desaturases / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008854:Microscopy, Electron; D008930:Mitochondria, Liver / Q000201:enzymology*; D009154:Mutation; D019301:Oleic Acid / Q000378:metabolism; D009829:Oleic Acids / Q000378:metabolism; D009842:Oligopeptides / Q000235:genetics; D044925:Oxidoreductases Acting on CH-CH Group Donors; D020675:Peroxisomes / Q000201:enzymology; D010957:Plasmids; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics
chemical_list: D005231:Fatty Acids, Unsaturated; D009829:Oleic Acids; D009842:Oligopeptides; C088449:seryl-lysyl-leucine; D019301:Oleic Acid; C008820:petroselinic acid; D044943:Fatty Acid Desaturases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C100153:2,4-dienoyl-CoA reductase; D019748:Carbon-Carbon Double Bond Isomerases; D064006:Dodecenoyl-CoA Isomerase; C576298:ECI1 protein, human; C119738:ECI2 protein, human