Crystallization and preliminary crystallographic data of 2-enoyl-CoA hydratase 2 domain of Candida tropicalis peroxisomal multifunctional enzyme type 2.
In yeast, the second and the third reaction of the fatty-acid beta-oxidation spiral are catalysed by peroxisomal multifunctional enzyme type 2 (Mfe2p/Fox2p). This protein has two (3R)-hydroxyacyl-CoA dehydrogenase domains and a C-terminal 2-enoyl-CoA hydratase 2 domain. Here, the purification, crystallization and X-ray diffraction analysis of the hydratase 2 domain [CtMfe2p(dh(a+b)Delta)] from Candida tropicalis Mfe2p is reported. CtMfe2p(dh(a+b)Delta) was overexpressed as an enzymatically active recombinant protein and crystallized by the hanging-drop vapour-diffusion method. The crystals belong to space group C2, with unit-cell parameters a = 178.57, b = 60.46, c = 130.85 A, beta = 94.48 degrees. Selenomethionine-labelled protein was used for a multi-wavelength anomalous dispersion (MAD) experiment. A three-wavelength data set suitable for MAD phasing was collected to 2.25 A resolution using synchrotron radiation.
Koski
M Kristian
MK
Department of Biochemistry, University of Oulu, Box 3000, FIN-90014 University of Oulu, Finland.
Haapalainen
Antti M
AM
Hiltunen
J Kalervo
JK
Glumoff
Tuomo
T
D041022:Candida tropicalis / Q000201:enzymology*; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray / Q000379:methods; D004746:Enoyl-CoA Hydratase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D005656:Fungal Proteins / Q000737:chemistry; D017434:Protein Structure, Tertiary; D012645:Selenomethionine
D005656:Fungal Proteins; D012645:Selenomethionine; D004746:Enoyl-CoA Hydratase; C083735:long-chain-enoyl-CoA hydratase