title: Cytochrome b₅ coexpression increases Tetrahymena thermophila Δ6 fatty acid desaturase activity in Saccharomyces cerevisiae.
abstract: Very-long-chain polyunsaturated fatty acids such as arachidonic, eicosapentaenoic, and docosahexaenoic acids, are important to the physiology of many microorganisms and metazoans and are vital to human development and health. The production of these and related fatty acids depends on Δ6 desaturases, the final components of an electron transfer chain that introduces double bonds into 18-carbon fatty acid chains. When a Δ6 desaturase identified from the ciliated protist Tetrahymena thermophila was expressed in Saccharomyces cerevisiae cultures supplemented with the 18:2(Δ9,12) substrate, only 4% of the incorporated substrate was desaturated. Cytochrome b₅ protein sequences identified from the genome of T. thermophila included one sequence with two conserved cytochrome b₅ domains. Desaturation by the Δ6 enzyme increased as much as 10-fold when T. thermophila cytochrome b₅s were coexpressed with the desaturase. Coexpression of a cytochrome b₅ from Arabidopsis thaliana with the Δ6 enzyme also increased desaturation. A split ubiquitin growth assay indicated that the strength of interaction between cytochrome b₅ proteins and the desaturase plays a vital role in fatty acid desaturase activity, illustrating the importance of protein-protein interactions in this enzyme activity.
authors.affiliation: Institute of Biological Chemistry Washington State University, Pullman, Washington, USA.
authors.forename: Jeremy L
authors.initials: JL
authors.lastname: Dahmen
authors.forename: Rebecca
authors.initials: R
authors.lastname: Olsen
authors.forename: Deirdre
authors.initials: D
authors.lastname: Fahy
authors.forename: James G
authors.initials: JG
authors.lastname: Wallis
authors.forename: John
authors.initials: J
authors.lastname: Browse
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D015786:Cytochromes b5 / Q000235:genetics* / Q000378:metabolism; D057075:Enzyme Assays; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D015870:Gene Expression; D050602:Linoleoyl-CoA Desaturase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D016808:Tetrahymena thermophila / Q000201:enzymology / Q000235:genetics*
chemical_list: D029681:Arabidopsis Proteins; D005231:Fatty Acids, Unsaturated; D015800:Protozoan Proteins; D015786:Cytochromes b5; D050602:Linoleoyl-CoA Desaturase