title: Crystallization and characterization of the dehydrogenase domain from rat peroxisomal multifunctional enzyme type 1.
abstract: Peroxisomal multifunctional enzyme type 1 from rat (perMFE-1) is a monomeric multidomain protein shown to have 2-enoyl-CoA hydratase/Delta(3)-Delta(2)-enoyl-CoA isomerase and (3S)-hydroxyacyl-CoA dehydrogenase domains followed by a C-terminal extension of 130 amino acids with unknown function apart from being a carrier of the peroxisomal targeting signal type 1. The truncated perMFE-1 without the N-terminal hydratase/isomerase domain (perMFE-1DH; residues 260-722) was overexpressed as an enzymatically active recombinant protein, purified and characterized. Using (3S)-hydroxydecanoyl-CoA as a substrate, the specific enzymatic activity of perMFE-1DH was determined to be 2.2 micromol min(-1) mg(-1), comparable with that of perMFE-1 purified from rat liver (2.8 micromol min(-1) mg(-1)). The protein was crystallized in the apo form by the hanging-drop method and a complete data set to 2.45 A resolution was collected using a rotating-anode X-ray source. The crystals have primitive tetragonal symmetry, with unit-cell parameters a = b = 125.9, c = 60.2 A.
authors.affiliation: Biocenter Oulu and Department of Biochemistry, University of Oulu, PO Box 3000, FIN-90014, Finland.
authors.forename: Jukka P
authors.initials: JP
authors.lastname: Taskinen
authors.forename: Tiila-Riikka
authors.initials: TR
authors.lastname: Kiema
authors.forename: Kari T
authors.initials: KT
authors.lastname: Koivuranta
authors.forename: Rik K
authors.initials: RK
authors.lastname: Wierenga
authors.forename: J Kalervo
authors.initials: JK
authors.lastname: Hiltunen
mesh_terms: D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000737:chemistry*; D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D004746:Enoyl-CoA Hydratase / Q000737:chemistry*; D007535:Isomerases / Q000737:chemistry*; D009097:Multienzyme Complexes / Q000737:chemistry*; D063994:Peroxisomal Bifunctional Enzyme; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D015094:3-Hydroxyacyl CoA Dehydrogenases; D004746:Enoyl-CoA Hydratase; D063994:Peroxisomal Bifunctional Enzyme; D007535:Isomerases