Liu Lili, O'Grady Christopher, Dalrymple Sean A, Prasad Lata, Dmitriev Oleg Y, Delbaere Louis T J
Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 5E5, Canada.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Jun 1;65(Pt 6):621-4. doi: 10.1107/S1744309109017023. Epub 2009 May 22.
Wilson disease associated protein (ATP7B) is essential for copper transport in human cells. Mutations that affect ATP7B function result in Wilson's disease, a chronic copper toxicosis. Disease-causing mutations within the N-domain of ATP7B (WND) are known to disrupt ATP binding, but a high-resolution X-ray structure of the ATP-binding site has not been reported. The N-domain was modified to delete the disordered loop comprising residues His1115-Asp1138 (WNDDelta(1115-1138)). Unlike the wild-type N-domain, WNDDelta(1115-1138) formed good-quality crystals. Synchrotron diffraction data have been collected from WNDDelta(1115-1138) at the Canadian Light Source. A native WNDDelta(1115-1138) crystal diffracted to 1.7 A resolution and belonged to space group P4(2)2(1)2, with unit-cell parameters a = 39.2, b = 39.2, c = 168.9 A. MAD data were collected to 2.7 A resolution from a SeMet-derivative crystal with unit-cell parameters a = 38.4, b = 38.4, c = 166.7 A. The WNDDelta(1115-1138) structure is likely to be solved by phasing from multiwavelength anomalous diffraction (MAD) experiments.
威尔逊病相关蛋白（ATP7B）对人类细胞中的铜转运至关重要。影响ATP7B功能的突变会导致威尔逊病，一种慢性铜中毒。已知ATP7B的N结构域（WND）内的致病突变会破坏ATP结合，但尚未报道ATP结合位点的高分辨率X射线结构。对N结构域进行了修饰，删除了包含His1115 - Asp1138残基的无序环（WNDDelta(1115 - 1138)）。与野生型N结构域不同，WNDDelta(1115 - 1138)形成了高质量的晶体。已在加拿大光源处从WNDDelta(1115 - 1138)收集了同步加速器衍射数据。一个天然的WNDDelta(1115 - 1138)晶体衍射到1.7 Å分辨率，属于空间群P4(2)2(1)2，晶胞参数a = 39.2，b = 39.2，c = 168.9 Å。从具有晶胞参数a = 38.4，b = 38.4，c = 166.7 Å的SeMet衍生物晶体收集了到2.7 Å分辨率的MAD数据。WNDDelta(1115 - 1138)结构可能通过多波长反常衍射（MAD）实验的相位确定来解析。
