STRUCTURAL STUDIES ON ADENOVIRUS CAPSIDS AND ASSOCIATED STRUCTURAL PROTEINS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 腺病毒是一种无包膜的DNA病毒，具有二十面体对称性，直径约950A。它们与急性呼吸道、胃肠道和眼部感染以及在基因传递应用中的应用引起了基础和医学研究人员的强烈兴趣。目前，美国约有28%的病毒载体基因治疗试验使用腺病毒，主要用于治疗心血管疾病和癌症。 由于顶端长而突出的纤维，很难使整个腺病毒结晶。我们最近结晶了一种形式的嵌合腺病毒，其衍射率超过5A分辨率。我们已经在APS的BIOCARS和GMCA光束线上收集了8A分辨率的部分数据集，并安装在毛细管上。最近，我们成功地确定了一种可以冻结腺病毒晶体的低温条件。我们想从冷冻晶体以及毛细管安装的晶体中收集完整的数据集。对腺病毒整个结构的了解提供了有关病毒的三维折叠和各种蛋白质之间的相互作用以及病毒的各种抗原区的信息。此外，该分子的大小约为150Mda，直径约为1000A，使其成为X射线结晶学解析的最大结构。在解决这一巨型结构中获得的知识促进了解决类似类型的大型结构所需的技术。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Adenoviruses (Ad) are non-enveloped DNA viruses with icosahedral symmetry and a diameter of approximately 950A. Their association with acute respiratory, gastrointestinal and ocular infections as well as use in gene delivery applications has drawn intense interest from basic and medical researchers. Approximately 28% of all viral vector gene therapy trials in the U.S. currently employ adenovirus, primarily for the treatment of cardiovascular diseases and cancer. It has been difficult to crystallize the entire adenovirus due to long protruding fibers from the vertices. We recently crystallized a form of chimeric adenovirus that diffracts beyond 5A resolution. We have collected partial dataset to 8A resolution at the BIOCARS and GMCA beam lines at APS on capillary mounted crystals. Recently we have successfully identified a cryo condition where the adenovirus crystals can be frozen. We would like to collect a complete data set from cryo frozen crystals as well as from capillary mounted crystals. Knowledge of the entire structure of adenovirus provides the information on the three dimensional fold and interactions between various proteins as well as various antigenic regions of the virus. Furthermore, the enormity of this molecule of the size of 150Mda and ~1000A in diameter make it a largest structure solved by the x-ray crystallography. Knowledge gained in solving this mega structure advances the technologies required for solving similar kinds of large structures.