Structure-function analysis of phytanoyl-CoA 2-hydroxylase mutations causing Refsum's disease.

Mukherji, M; Chien, W; Kershaw, N; Clifton, I; Schofield, C; Wierzbicki, A; Lloyd, MD

Journal article

Structure-function analysis of phytanoyl-CoA 2-hydroxylase mutations causing Refsum's disease.

Abstract:: Refsum's disease is a neurological syndrome characterized by adult-onset retinitis pigmentosa, anosmia, sensory neuropathy and phytanic acidaemia. Many cases are caused by mutations in peroxisomal oxygenase phytanoyl-CoA 2-hydroxylase (PAHX) which catalyses the initial alpha-oxidation step in the degradation of phytanic acid. Both pro and mature forms of recombinant PAHX were produced in Escherichia coli, highly purified, and shown to have a requirement for iron(II) as a co-factor and 2-oxoglutarate as a co-substrate. Sequence analysis in the light of crystallographic data for other members of the 2-oxoglutarate-dependent oxygenase super-family led to secondary structural predictions for PAHX, which were tested by site-directed mutagenesis. The H175A and D177A mutants did not catalyse hydroxylation of phytanoyl-CoA, consistent with their assigned role as iron(II) binding ligands. The clinically observed P29S, Q176K, G204S, N269H, R275Q and R275W mutants were assayed for both 2-oxoglutarate and phytanoyl-CoA oxidation. The P29S mutant was fully active, implying that the mutation resulted in defective targeting of the protein to peroxisomes. Mutation of Arg-275 resulted in impaired 2-oxoglutarate binding. The Q176K, G204S and N269H mutations caused partial uncoupling of 2-oxoglutarate conversion from phytanoyl-CoA oxidation. The results demonstrate that the diagnosis of Refsum's disease should not solely rely upon PAHX assays for 2-oxoglutarate or phytanoyl-CoA oxidation.

Publication status:: Published

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APA Style

Mukherji, M., Chien, W., Kershaw, N., Clifton, I., Schofield, C., Wierzbicki, A., & Lloyd, M. D. (2001). Structure-function analysis of phytanoyl-CoA 2-hydroxylase mutations causing Refsum's disease. Human Molecular Genetics, 10(18), 1971–1982.

MLA Style

Mukherji, M, et al. “Structure-Function Analysis of Phytanoyl-CoA 2-Hydroxylase Mutations Causing Refsum's Disease.” Human Molecular Genetics, vol. 10, no. 18, 2001, pp. 1971–82.

Chicago Style

Mukherji, M, W Chien, N Kershaw, et al. 2001. “Structure-Function Analysis of Phytanoyl-CoA 2-Hydroxylase Mutations Causing Refsum's Disease.” Human Molecular Genetics 10 (18): 1971–82.
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