Crystallization and preliminary X-ray crystallographic analysis of eIF5BN and the eIF5BΔN-eIF1AΔN complex

Abstract

The binding between two universally conserved translation initiation factors, eIF5B and eIF1A, is important in the initiation step of eukaryotic protein synthesis on the ribosome. Through this interaction, eIF1A assists in recruiting eIF5B to the initiating 40S subunit; eIF5B then encourages the joining of the 60S subunit to form an initiating 80S ribosome. Here, the expression, purification, crystallization and preliminary X-ray analyses of eIF5BN and the eIF5BN-eIF1AN complex from Saccharomyces cerevisiae are reported. The crystal of eIF5BN diffracted to 2.45 Å resolution and belonged to space group P41212, with unit-cell parameters a = b = 130.0, c = 71.7 Å. The asymmetric unit was estimated to contain one molecule. The initial phase was obtained by Se-SAD. The crystal of the eIF5BN-eIF1AN complex diffracted to 3.3 Å resolution and belonged to space group P212121, with unit-cell parameters a = 101.9, b = 120.9, c = 132.8 Å. The asymmetric unit was estimated to contain two complex molecules. © 2011 International Union of Crystallography. All rights reserved.