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Title: Role of thiamine pyrophosphate in oligomerisation, functioning and import of peroxisomal 2-hydroxyacyl-CoA lyase
Authors: Fraccascia, Patrizia
Casteels, Minne
De Schryver, Evelyn
Van Veldhoven, Paul P # ×
Issue Date: Oct-2011
Publisher: Elsevier
Series Title: Biochimica et Biophysica Acta. Proteins and Proteomics vol:1814 issue:10 pages:1226-1233
Abstract: During peroxisomal α-oxidation, the CoA-esters of phytanic acid and 2-hydroxylated straight chain fatty acids are cleaved into a (n-1) fatty aldehyde and formyl-CoA by 2-hydroxyacyl-CoA lyase (HACL1). HACL1 is imported into peroxisomes via the PEX5/PTS1 pathway, and so far, it is the only known peroxisomal TPP-dependent enzyme in mammals. In this study, the effect of mutations in the TPP-binding domain of HACL1 on enzyme activity, subcellular localisation and oligomerisation was investigated. Mutations of the aspartate 455 and serine 456 residues within the TPP binding domain of the human HACL1 did not affect the targeting upon expression in transfected CHO cells, although enzyme activity was abolished. Gel filtration of native and mutated N-His(6)-fusions, expressed in yeast, revealed that the mutations did not influence the oligomerisation of the (apo)enzyme. Subcellular fractionation of yeast cells expressing HACL1 showed that the lyase activity sedimented at high density in a Nycodenz gradient. In these fractions TPP could be measured, but not when mutated HACL1 was expressed, although the recombinant enzyme was still targeted to peroxisomes. These findings indicate that the binding of TPP is not required for peroxisomal targeting and correct folding of HACL1, in contrast to other TPP-dependent enzymes, and suggest that transport of TPP into peroxisomes is dependent on HACL1 import, without requirement of a specific solute transporter.
URI: 
ISSN: 1570-9639
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Department of Pharmaceutical & Pharmacological Sciences - miscellaneous
Laboratory of Lipid Biochemistry and Protein Interactions
Pharmacology Section (-)
× corresponding author
# (joint) last author

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