RRC ID 586
Author Pedersen L, Henriksen A.
Title Expression, purification and crystallization of two peroxisomal acyl-CoA oxidases from Arabidopsis thaliana.
Journal Acta Crystallogr D Biol Crystallogr
Abstract Two members of the acyl-CoA oxidase family from Arabidopsis thaliana have been cloned, overexpressed, purified and crystallized. Long-chain-specific acyl-CoA oxidase 1 crystals are characterized by a large variation in diffraction quality and non-isomorphous unit-cell parameters. The best crystals diffract to 2.0 angstrom using synchrotron radiation, have unit-cell parameters a = 85.2, b = 117.0, c = 131.0 angstrom, alpha = beta = gamma = 90 degrees and show P2(1)2(1)2(1) symmetry. There are two polypeptide chains in the asymmetric unit. Short-chain-specific acyl-CoA oxidase 4 crystals are trigonal, space group P3(1)21/P3(2)21, with unit-cell parameters a = b = 198.7, c = 149.6 angstroms. The crystals are most likely to contain six polypeptide chains in the asymmetric unit. Freshly prepared acyl-CoA oxidase 4 crystals diffract to 3.9 angstroms at cryogenic temperature at beamline I711, Max-Lab, but the diffraction quality degenerates after storage for only a few days in the crystallization drop. A selenomethionine-substituted form of the protein was produced and two-wavelength MAD data were collected at beamline BW7A, EMBL Outstation, Hamburg.
Volume 60(Pt 6)
Pages 1125-8
Published 2004-6-1
DOI 10.1107/S0907444904007577
PII S0907444904007577
PMID 15159576
MeSH Acyl-CoA Oxidase / chemistry* Acyl-CoA Oxidase / genetics Arabidopsis / enzymology* Arabidopsis Proteins / chemistry* Arabidopsis Proteins / genetics Cloning, Molecular Crystallography, X-Ray / methods* Genes, Plant Models, Chemical Peptides / chemistry Peroxisomes / enzymology* Plasmids / metabolism Synchrotrons Temperature Time Factors X-Ray Diffraction
IF 3.227
Times Cited 6
Arabidopsis / Cultured plant cells, genes pda05283