Studies on the specificity of unprocessed and mature forms of phytanoyl-CoA 2-hydroxylase and mutation of the iron binding ligands

The mature form of phytanoyl-coenzyme A 2-hydroxylase (PAHX), a nonheme Fe(II)- and 2-oxoglutarate-dependent oxygenase, catalyzes the α-hydroxylation of phytanoyl-CoA within peroxisomes. Mutations in PAHX result in some forms of adult Refsum's disease. Unprocessed PAHX (pro-PAHX) contains an N-...

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Main Authors:	Timothy Searls, Danica Butler, Winnie Chien, Mridul Mukherji, Matthew D. Lloyd, Christopher J. Schofield
Format:	Article
Language:	English
Published:	Elsevier 2005-08-01
Series:	Journal of Lipid Research
Subjects:	chemical rescue oxygenase 2-oxoglutarate phytanoyl-coenzyme A 2-hydroxylase phytanic acid Refsum's disease
Online Access:	http://www.sciencedirect.com/science/article/pii/S0022227520329655

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spelling	doaj-60cef5c4d0704254935a7959317f6b022021-04-27T04:43:51ZengElsevierJournal of Lipid Research0022-22752005-08-0146816601667Studies on the specificity of unprocessed and mature forms of phytanoyl-CoA 2-hydroxylase and mutation of the iron binding ligandsTimothy Searls0Danica Butler1Winnie Chien2Mridul Mukherji3Matthew D. Lloyd4Christopher J. Schofield5Chemistry Research Laboratory, University of Oxford, Oxford OX1 3TA, UKChemistry Research Laboratory, University of Oxford, Oxford OX1 3TA, UKChemistry Research Laboratory, University of Oxford, Oxford OX1 3TA, UKChemistry Research Laboratory, University of Oxford, Oxford OX1 3TA, UKDepartment of Pharmacy and Pharmacology, University of Bath, Bath BA2 7AY, UKTo whom correspondence should be addressed.; Chemistry Research Laboratory, University of Oxford, Oxford OX1 3TA, UKThe mature form of phytanoyl-coenzyme A 2-hydroxylase (PAHX), a nonheme Fe(II)- and 2-oxoglutarate-dependent oxygenase, catalyzes the α-hydroxylation of phytanoyl-CoA within peroxisomes. Mutations in PAHX result in some forms of adult Refsum's disease. Unprocessed PAHX (pro-PAHX) contains an N-terminal peroxisomal targeting sequence that is cleaved to give mature PAHX (mat-PAHX). Previous studies have implied a difference in the substrate specificity of the unprocessed and mature forms of PAHX. We demonstrate that both forms are able to hydroxylate a range of CoA derivatives, but under the same assay conditions, the N-terminal hexa-His-tagged unprocessed form is less active than the nontagged mature form. Analyses of the assay conditions suggest a rationale for the lack of activity previously reported for some substrates (e.g. isovaleryl-CoA) for the (His)6pro-PAHX.Site-directed mutagenesis was used to support proposals for the identity of the iron binding ligands (His-175, Asp-177, His-264) of the 2-His-1-carboxylate motif of PAHX. Mutation of other histidine residues (His-213, His-220, His-259) suggested that these residues were not involved in Fe(II) binding.http://www.sciencedirect.com/science/article/pii/S0022227520329655chemical rescueoxygenase2-oxoglutaratephytanoyl-coenzyme A 2-hydroxylasephytanic acidRefsum's disease
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Timothy Searls Danica Butler Winnie Chien Mridul Mukherji Matthew D. Lloyd Christopher J. Schofield
spellingShingle	Timothy Searls Danica Butler Winnie Chien Mridul Mukherji Matthew D. Lloyd Christopher J. Schofield Studies on the specificity of unprocessed and mature forms of phytanoyl-CoA 2-hydroxylase and mutation of the iron binding ligands Journal of Lipid Research chemical rescue oxygenase 2-oxoglutarate phytanoyl-coenzyme A 2-hydroxylase phytanic acid Refsum's disease
author_facet	Timothy Searls Danica Butler Winnie Chien Mridul Mukherji Matthew D. Lloyd Christopher J. Schofield
author_sort	Timothy Searls
title	Studies on the specificity of unprocessed and mature forms of phytanoyl-CoA 2-hydroxylase and mutation of the iron binding ligands
title_short	Studies on the specificity of unprocessed and mature forms of phytanoyl-CoA 2-hydroxylase and mutation of the iron binding ligands
title_full	Studies on the specificity of unprocessed and mature forms of phytanoyl-CoA 2-hydroxylase and mutation of the iron binding ligands
title_fullStr	Studies on the specificity of unprocessed and mature forms of phytanoyl-CoA 2-hydroxylase and mutation of the iron binding ligands
title_full_unstemmed	Studies on the specificity of unprocessed and mature forms of phytanoyl-CoA 2-hydroxylase and mutation of the iron binding ligands
title_sort	studies on the specificity of unprocessed and mature forms of phytanoyl-coa 2-hydroxylase and mutation of the iron binding ligands
publisher	Elsevier
series	Journal of Lipid Research
issn	0022-2275
publishDate	2005-08-01
description	The mature form of phytanoyl-coenzyme A 2-hydroxylase (PAHX), a nonheme Fe(II)- and 2-oxoglutarate-dependent oxygenase, catalyzes the α-hydroxylation of phytanoyl-CoA within peroxisomes. Mutations in PAHX result in some forms of adult Refsum's disease. Unprocessed PAHX (pro-PAHX) contains an N-terminal peroxisomal targeting sequence that is cleaved to give mature PAHX (mat-PAHX). Previous studies have implied a difference in the substrate specificity of the unprocessed and mature forms of PAHX. We demonstrate that both forms are able to hydroxylate a range of CoA derivatives, but under the same assay conditions, the N-terminal hexa-His-tagged unprocessed form is less active than the nontagged mature form. Analyses of the assay conditions suggest a rationale for the lack of activity previously reported for some substrates (e.g. isovaleryl-CoA) for the (His)6pro-PAHX.Site-directed mutagenesis was used to support proposals for the identity of the iron binding ligands (His-175, Asp-177, His-264) of the 2-His-1-carboxylate motif of PAHX. Mutation of other histidine residues (His-213, His-220, His-259) suggested that these residues were not involved in Fe(II) binding.
topic	chemical rescue oxygenase 2-oxoglutarate phytanoyl-coenzyme A 2-hydroxylase phytanic acid Refsum's disease
url	http://www.sciencedirect.com/science/article/pii/S0022227520329655
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Studies on the specificity of unprocessed and mature forms of phytanoyl-CoA 2-hydroxylase and mutation of the iron binding ligands

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