CRYSTALLIZATION AND PRELIMINARY-X-RAY DIFFRACTION STUDY OF RECOMBINANT HUMAN EUKARYOTIC INITIATION FACTOR-4E

Recombinant human eukaryotic initiation factor-4E (eIF-4E), purified b y m(7)GTP-Sepha-rose 4B affinity chromatography, was used for crystall ization. After concentration of the eIF-4E protein (7 mg/ml), the solu tion was subjected to crystallization by the hanging-drop method. Tran sparent needle crystals complexed with m(7)GTP were obtained from 50 m M 2-(N-morpholino)ethanesulfonic acid-KOH buffer (pH 6.5) containing 2 5% (w/v) polyethylene glycol 6000 and 0.2 M (NH4)(2)SO4. The crystals belong to tetragonal space group P4(1) or P4(3), of Z=4, with unit-cel l dimensions of a=89.26, b=89.26, and c=38.51 Angstrom, and diffract b eyond 2.1 Angstrom resolution. The V-m value was calculated to be 3.07 Angstrom(3)/Da, which indicates a solvent content of 59.9%.