II型单纯疱疹病毒糖蛋白D的晶体结构解析

X-ray diffraction analysis of glycoprotein D from Herpes simplex virus type 2

摘要:

II型单纯疱疹病毒 (HSV-2) 糖蛋白D (Glycoprotein D，gD) 在介导该病毒入侵到宿主细胞中起着关键作用。为了更好地研究gD在病毒侵入过程中的作用机制，利用杆状病毒表达系统表达了gD胞外部分区域 (1~285 aa)，通过Ni-NTA亲和层析以及分子排阻层析纯化后，得到的带His标签的分泌型可溶蛋白，用悬滴法对该蛋白进行了晶体筛选，获得了高质量的晶体。晶体生长条件为0.1 mol/L Hepes缓冲液 (pH 7.2)，5％ (V/V) 2-甲基-2,4-戊二醇 (MPD)，10％

Abstract:

Glycoprotein D (gD) of Herpes simplex virus type 2 (HSV-2) is a key factor mediating the entry of HSV-2 into host cells. In order to explain the mechanism underlying the gD-mediated receptor-binding and viral entry, we performed a structural study on HSV-2 gD. The ectodomain of the gD protein encompassing residues 1 to 285 was expressed by baculovirus-infected insect cells as a secreted soluble protein with a C-terminal hexa-his tag. The protein was then purified by affinity and size-exclusion chromatography. The purified protein was successfully crystallized using the hanging-drop vapor-diffusion at 18 °C in a condition consisting of 0.1 mol/L Hepes pH 7.2, 5% (V/V) 2-methyl-2, 4-pentanediol (MPD) and 10% PEG 10 000. The crystals diffracted to 1.8 ? resolution and belonged to space group P21, with unit-cell parameters a=63.6, b=55.4, c=65.3 ?, β=96.3°.