STRUCTURAL BIOLOGY OF MACROMOLECULAR STRUCTURE

本实验室旨在阐明控制 大分子的折叠和超分子的组装 复合物，并作为细胞器、细胞和组织合成的基础。 为此，基于电子的结构分析的先进方法 显微技术和计算机图像处理技术得到了发展和应用。 在 在过去的一年里，我们应用了冷冻电子显微镜和三个- 单纯疱疹病毒纯化衣壳的三维图像重建 病毒，1型，已在体外生化耗尽的某些 组件，与这些组件重新关联，或用 克隆抗体 结果导致了临时的 定位的四个小衣壳蛋白，并有影响 他们可能的功能角色。 VP 22（40 kDa）完全位于 的壳体内 （前体）B-衣壳，但当DNA 被包装：最有可能的是，它是一种支架蛋白， 部分地负责控制大约 900个拷贝的主要衣壳蛋白-VP 5（148 kDa）-进入一个大的 （125 nm），精确定义，二十面体壳。 VP 26（12 kDa）似乎 组成五邻体，其中存在80-100个这种小蛋白质的拷贝， 五重顶点中的每一个：VP 26可以参与控制 释放包装好的DNA VP 19（55 kDa）和VP 23（36 kDa）似乎 是三链体蛋白，在三重位点形成异源三聚体 在二十面体表面晶格的外表面上。 通过类比 与其他病毒的类似蛋白质，它们可能有助于 通过加强下面的阵列来稳定衣壳壳， VP 5的六聚体。

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.