AD2 PROTEINASE--TARGET FOR ANTIVIRAL THERAPY

在许多抗病毒治疗的靶点中， 某些病毒感染是病毒编码的蛋白酶。这些 需要酶来处理所涉及的病毒特异性前体 在这种致病菌的成熟、组装和复制中 人类病毒如脊髓灰质炎病毒、脑炎病毒、乙肝病毒、 和人类免疫缺陷病毒。这些病毒编码的 蛋白水解酶对其病毒编码底物具有高度的特异性。 因此，如果能够开发出同样特定的抑制剂并进行靶向 对于受感染的细胞，它们应该干扰病毒的复制和 没有正常的细胞新陈代谢。 人腺病毒2编码一种可处理6个病毒粒子的蛋白酶 病毒形态发生过程中的多肽。在缺少 活性病毒编码的蛋白水解酶，产生非传染性病毒。 因为人们对腺病毒的分子生物学了解很多。 2，由于其病毒编码的蛋白酶是一种丝氨酸蛋白酶， 腺病毒2是一个很好的模型系统来测试腺病毒的疗效 作为抗病毒药物的蛋白酶抑制剂。四大中的 蛋白水解酶、丝氨酸蛋白酶及其抑制物的种类如下 到目前为止最具特点的。 AD2蛋白酶将被克隆、表达、纯化和 描述一下。牛胰蛋白酶抑制因子的突变体Will 合理而随机地设计、克隆、表达和 被选为AD2蛋白水解酶的特异性抑制剂。牛 选择胰蛋白酶抑制剂是因为它只有58 氨基酸，对类胰酶丝氨酸蛋白酶有很高的亲和力， 并已被克隆并在大肠杆菌中以活性形式表达。 在选择了其他特性后，突变的牛 将测试胰腺胰蛋白酶抑制剂的抗病毒活性 腺病毒2型模型系统。如果成功，吸取的教训 从这个模型系统可以很容易地扩展到更多的医学上 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.