le Maire Albane, Schiltz Marc, Braud Sandrine, Gondry Muriel, Charbonnier Jean-Baptiste, Zinn-Justin Sophie, Stura Enrico
Département d'Ingénierie et d'Etude des Protéines, Commissariat a l'Energie Atomique, 91191 Gif-sur-Yvette, France.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Mar 1;62(Pt 3):245-8. doi: 10.1107/S174430910600409X. Epub 2006 Feb 10.
Here, the crystallization and initial phasing of the C-terminal domain of human KIN17, a 45 kDa protein mainly expressed in response to ionizing radiation and overexpressed in certain tumour cell lines, are reported. Crystals diffracting to 1.4 A resolution were obtained from 10% ethylene glycol, 27% PEG 6000, 500 mM LiCl and 100 mM sodium acetate pH 6.3 in space group P2(1)2(1)2(1), with unit-cell parameters a = 45.75, b = 46.31, c = 60.80 A and one molecule in the asymmetric unit. Since this domain has a basic pI, heavy-atom derivatives were obtained by soaking the crystals with negatively charged ions such as tungstate and iodine. The replacement of LiCl by KI in the cryosolution allowed the determination of phases from iodide ions to give an interpretable electron-density map.
本文报道了人KIN17蛋白C端结构域的结晶及初步相位分析。KIN17是一种45 kDa的蛋白,主要在受到电离辐射时表达,并在某些肿瘤细胞系中过表达。通过在含有10%乙二醇、27%聚乙二醇6000、500 mM氯化锂和100 mM醋酸钠(pH 6.3)的体系中培养,获得了分辨率为1.4 Å的晶体,其空间群为P2(1)2(1)2(1),晶胞参数a = 45.75、b = 46.31、c = 60.80 Å,不对称单元中有一个分子。由于该结构域具有碱性pI值,通过用带负电荷的离子(如钨酸盐和碘)浸泡晶体获得了重原子衍生物。在冷冻溶液中用碘化钾替代氯化锂,使得能够从碘离子确定相位,从而得到可解释的电子密度图。