AD2 PROTEINASE--TARGET FOR ANTIVIRAL THERAPY

在抗病毒治疗的许多目标中， 某些病毒感染是病毒编码的蛋白酶。 这些 需要酶来处理病毒特异性前体， 在这种致病性的成熟、组装和复制中， 人病毒如脊髓灰质炎病毒、脑炎病毒、B肝炎病毒， 和人类免疫缺陷病毒。 这些病毒编码的 蛋白酶对其病毒编码的底物具有高度特异性。 因此，如果同样特异性的抑制剂可以开发和靶向 感染的细胞，他们应该干扰病毒复制， 而不是正常的细胞代谢。 人腺病毒2编码一种蛋白酶，可处理六种病毒粒子 多肽在病毒形态发生过程中的作用。 在没有 活性病毒编码的蛋白酶，产生非感染性病毒。 因为我们对腺病毒的分子生物学了解很多 2，并且由于其病毒编码的蛋白酶是丝氨酸蛋白酶， 腺病毒2是一个很好的模型系统，以测试的效力， 蛋白酶抑制剂作为抗病毒剂。 四大 蛋白酶、丝氨酸蛋白酶及其抑制剂的种类， 到目前为止最好的特点。 Ad2蛋白酶将被克隆、表达、纯化和表达。 描述。 牛胰蛋白酶抑制剂的突变体将 合理随机设计、克隆、表达， 选择作为Ad2蛋白酶的特异性抑制剂。 牛 选择胰蛋白酶抑制剂，因为它只有58 氨基酸，对胰蛋白酶样丝氨酸蛋白酶具有高亲和力， 并已在E.杆菌 在选择额外的特性后， 胰胰蛋白酶抑制剂将被测试的抗病毒活性， 腺病毒2模型系统。 如果成功， 从这个模型系统可以很容易地扩展到更多的医学 相关病毒

Among the many targets for antiviral therapy that arise during certain viral infections are the virus-coded proteinases. These enzymes are required to process virus-specific precursors involved in the maturation, assembly and replication of such pathogenic human viruses as poliovirus, encephalitis virus, hepatitus B virus, and human immunodeficiency virus. These virus-coded proteinases are highly specific for their virus-coded substrates. Thus, if equally specific inhibitors can be developed and targeted to infected cells, they should interfere with virus replication and not with normal cellular metabolism. Human adenovirus 2 encodes a proteinase that processes six virion polypeptides during virus morphogenesis. In the absence of an active virus-coded proteinase, noninfectious virus is produced. Because much is known about the molecular biology of adenovirus 2, and because its virus-coded proteinase is a serine proteinase, adenovirus 2 is a good model system to test the efficacy of proteinase inhibitors as antiviral agents. Of the four major classes of proteinases, serine proteinases and their inhibitors are by far the best characterized. The Ad2 proteinase will be cloned, expressed, purified and characterize. Mutants of bovine pancreatic trypsin inhibitor will be rationally and randomly designed, cloned, expressed and selected as specific inhibitors of the Ad2 proteinase. Bovine pancreatic trypsin inhibitor is chosen, because it has only 58 amino acids, has a high affinity for trypsin-like serine proteinases, and has been cloned and expressed in an active form in E. coli. After selecting for additional properties, mutant bovine pancreatic trypsin inhibitors will be tested for antiviral activity in the adenovirus 2 model system. If successful, the lessons learned from this model system can easily be extended to more medically relevant viruses.