Open AccessPreliminary crystallographic analysis of the N‐terminal domain of FILIA, a protein essential for embryogenesis
Author(s) -
Wang Juke,
Zhang TongCun,
Liu Xinqi
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110031994
Subject(s) - orthorhombic crystal system , crystallography , crystallization , tetragonal crystal system , ammonium sulfate , molecule , diffraction , crystal (programming language) , crystal structure , chemistry , materials science , chromatography , organic chemistry , programming language , physics , computer science , optics
FILIA is a component of the subcortical maternal complex that is essential for early stage embryogenesis. Its 6×His‐tagged N‐terminal domain was expressed in Escherichia coli and purified to homogeneity. Two types of crystals formed under different crystallization conditions during screening. Orthorhombic crystals appeared in a solution containing 1.4 M ammonium sulfate, 0.1 M Tris pH 8.2 and 12% glycerol, while tetragonal crystals were obtained using 15% PEG 4000 mixed with 0.1 M HEPES pH 7.5 and 15% 2‐propanol. High‐quality diffraction data were collected from the two crystal forms to resolutions of 1.8 and 2.2 Å, respectively, using synchrotron radiation. The Matthews coefficients indicated that the P 2 1 2 1 2 1 and P 4 1 2 1 2 crystals contained two molecules and one molecule per asymmetric unit, respectively. A selenomethionine‐substituted sample failed to crystallize under the native conditions, but another orthorhombic crystal form was obtained under different conditions and anomalous diffraction data were collected.