STRUCTURAL BIOLOGY OF MACROMOLECULAR STRUCTURE

这个实验室的目的是阐明 大分子的折叠和超分子的组装 复合体，是细胞器、细胞和组织合成的基础。 为此，先进的基于电子的结构分析方法 显微镜和计算机图像处理技术得到了发展和应用。在……里面 在过去的一年里，我们应用了冷冻电子显微镜和三个- 纯化单纯疱疹病毒衣壳的三维图像重建 病毒，1型，在体外已经被生化耗尽的某些 组件，与这些组件重新关联，或用 单克隆抗体。结果导致了临时的 四种微小衣壳蛋白的定位及其意义 他们可能扮演的职能角色。VP22(40 KDa)完全位于 在(前体)B衣壳内，但当DNA 被包装：最有可能的是，它是一种支架蛋白，至少 部分负责控制聚合，约 900个拷贝的主要衣壳蛋白-VP5(148 KDa)-进入一个大的 (125 Nm)，精确定义的二十面体外壳。VP26(12 KDa)似乎 组成五角体，这种小蛋白质的80-100个拷贝存在于 五重顶点中的每一个：VP26可能参与控制 释放包装好的DNA。VP19(55 KDa)和VP23(36 KDa)似乎 是三链蛋白，在三个位点形成异三聚体 在二十面体表面晶格的外表面上。以此类推 与其他病毒的类似蛋白质一样，它们可能起到 通过加强底层的阵列来稳定衣壳 VP5的六角体。

This Laboratory aims to elucidate the regulatory mechanisms that govern the folding of macromolecules and the assembly of supramolecular complexes, and underlie the synthesis of organelles, cells, and tissues. To this end, advanced methods of structural analysis based on electron microscopy and computer image processing are developed and applied. In the past year, we have applied cryoelectron microscopy and three- dimensional image reconstruction to purified capsids of herpes simplex virus, type 1, which had been biochemically depleted in vitro of certain components, reassociated with those components, or decorated with monoclonal antibodies. The results have led to a provisional localization of the four minor capsid proteins, and have implications for their likely functional roles. VP22 (40kDa) is located entirely within the shell of (precursor) B-capsids, but is eliminated when DNA is packaged: most likely, it is a scaffolding protein that is at least partly responsible for controlling the polymerization of approximately 900 copies of the major capsid protein - VP5 (148kDa) - into a large (125nm), precisely defined, icosahedral shell. VP26 (12kDa) appears to make up the pentons, with 80-100 copies of this small protein present at each of the five-fold vertices: VP26 may be involved in controlling release of the packaged DNA. VP19 (55 kDa) and VP23 (36 kDa) appear to be the triplex proteins, forming heterotrimers at the three-fold sites on the outer surface of the icosahedral surface lattice. By analogy with comparable proteins of other viruses, they probably serve to stabilize the capsid shell by reinforcing the underlying array of hexamers of VP5.