Phytanoyl-CoA hydroxylase: recognition of 3-methyl-branched acyl-CoAs and requirement for GTP or ATP and Mg2+ in addition to its known hydroxylation cofactors (original) (raw)

2000, Journal of Lipid Research

Phytanoyl-CoA hydroxylase is a peroxisomal ␣oxidation enzyme that catalyzes the 2-hydroxylation of 3methyl-branched acyl-CoAs. A polyhistidine-tagged human phytanoyl-CoA hydroxylase was expressed in E. coli and subsequently purified as an active protein. The recombinant enzyme required GTP or ATP and Mg 2 ؉ , in addition to its known cofactors Fe 2 ؉ , 2-oxoglutarate, and ascorbate. The enzyme was active towards phytanoyl-CoA and 3-methylhexadecanoyl-CoA, but not towards 3-methylhexadecanoic acid. Racemic, R -and S -3-methylhexadecanoyl-CoA were equally well hydroxylated. Hydroxylation of R -and S -3methylhexadecanoyl-CoA yielded the ( 2S,3R ) and ( 2R,3S ) isomers of 2-hydroxy-3-methylhexadecanoyl-CoA, respectively. Human phytanoyl-CoA hydroxylase did not show any activity towards 2-methyl-and 4-methyl-branched acyl-CoAs or towards long and very long straight chain acyl-CoAs, excluding a possible role for the enzyme in the formation of 2hydroxylated and odd-numbered straight chain fatty acids, which are abundantly present in brain. In conclusion, we report the unexpected requirement for ATP or GTP and Mg 2 ؉ of phytanoyl-CoA hydroxylase in addition to the known hydroxylation cofactors. Due to the fact that straight chain fatty acyl-CoAs are not a substrate for phytanoyl-CoA hydroxylase, 2-hydroxylation of fatty acids in brain can be allocated to a different enzyme/pathway. -Croes, K., V. Foulon, M. Casteels, P. P. Van Veldhoven, and G. P. Mannaerts. Phytanoyl-CoA hydroxylase: recognition of 3-methylbranched acyl-CoAs and requirement for GTP or ATP and Mg 2 ؉ in addition to its known hydroxylation cofactors. J. Lipid Res. 2000. 41: 629-636.