Seto Azusa, Ikushima Hiroaki, Suzuki Toshiyasu, Sato Yusuke, Fukai Shuya, Yuki Keiko, Miyazawa Keiji, Miyazono Kohei, Ishitani Ryuichiro, Nureki Osamu
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama-shi, Kanagawa, Japan.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Jan 1;65(Pt 1):21-4. doi: 10.1107/S1744309108038219. Epub 2008 Dec 25.
GCIP/HHM is a human nuclear protein that is implicated in regulation of cell proliferation. Its primary structure contains helix-loop-helix and leucine-zipper motifs but lacks a DNA-binding basic region. Native and selenomethionine-derivatized (SeMet) crystals of full-length GCIP/HHM were obtained using the hanging-drop vapour-diffusion method. The crystals were greatly improved by adding tris(2-carboxyethyl)phosphine as a reducing reagent and diffracted to 3.5 A resolution. Preliminary phase calculations using the data set obtained from the SeMet crystal suggested that the crystal belonged to space group P3(2)21 and contained one molecule per asymmetric unit. Structure determination by the multiple-wavelength anomalous dispersion method using the SeMet crystals is in progress.
GCIP/HHM是一种涉及细胞增殖调控的人类核蛋白。其一级结构包含螺旋-环-螺旋和亮氨酸拉链基序，但缺乏DNA结合碱性区域。使用悬滴气相扩散法获得了全长GCIP/HHM的天然晶体和硒代甲硫氨酸衍生化（SeMet）晶体。通过添加三（2-羧乙基）膦作为还原剂，晶体得到了极大改善，衍射分辨率达到3.5埃。使用从SeMet晶体获得的数据集进行的初步相位计算表明，该晶体属于空间群P3(2)21，每个不对称单元包含一个分子。使用SeMet晶体通过多波长反常色散法进行结构测定正在进行中。
