title: Crystallization and preliminary X-ray diffraction study of recombinant human eukaryotic initiation factor-4E.
abstract: Recombinant human eukaryotic initiation factor-4E (eIF-4E), purified by m7GTP-Sepharose 4B affinity chromatography, was used for crystallization. After concentration of the eIF-4E protein (7 mg/ml), the solution was subjected to crystallization by the hanging-drop method. Transparent needle crystals complexed with m7GTP were obtained from 50 mM 2-(N-morpholino)ethanesulfonic acid-KOH buffer (pH 6.5) containing 25% (w/v) polyethylene glycol 6000 and 0.2 M (NH4)2SO4. The crystals belong to tetragonal space group P4(1) or P4(3), of Z = 4, with unit-cell dimensions of a = 89.26, b = 89.26, and c = 38.51 angstrum, and diffract beyond 2.1 angstrum resolution. The Vm value was calculated to be 3.07 angstrum 3/Da, which indicates a solvent content of 59.9%.
authors.affiliation: Department of Physical Chemistry, Osaka University of Pharmaceutical Sciences.
authors.forename: S
authors.initials: S
authors.lastname: Morino
authors.forename: K
authors.initials: K
authors.lastname: Tomoo
authors.forename: N
authors.initials: N
authors.lastname: Nishi
authors.forename: K
authors.initials: K
authors.lastname: Okabe
authors.forename: M
authors.initials: M
authors.lastname: Doi
authors.forename: T
authors.initials: T
authors.lastname: Ishida
authors.forename: K
authors.initials: K
authors.lastname: Kitamura
mesh_terms: D003460:Crystallization; D039561:Eukaryotic Initiation Factor-4E; D006801:Humans; D010448:Peptide Initiation Factors / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D014961:X-Ray Diffraction
chemical_list: D039561:Eukaryotic Initiation Factor-4E; D010448:Peptide Initiation Factors; D011994:Recombinant Proteins