PICORNAVIRAL PROCESSING PROTEINASES

我们的长期目标是发展强有力的和有选择性的 小核糖核酸病毒加工酶的抑制剂，可能在 抗病毒治疗。 为了实现这一目标，我们将研究催化和结合 甲型肝炎病毒3C型半胱氨酸蛋白酶的性质 鼻病毒2型和14型以及人类脊髓灰质炎病毒。合作小组， 由Protos的Bruce Malcolm领导的，表达了大量的 肝炎酵素。我们已经研究了各种潜在的多肽亚组分 并开发了快速、定量的分析方法。我们现在准备好了 通过改进识别的多肽结构来扩大我们的研究范围 这种酶，通过开发基于敏感的新的检测方法 荧光方法，并通过构建潜在的抑制剂基于 多肽结构。我们将利用所有这些方法来彻底 分析底物和抑制剂与肝炎的相互作用 酵素。 由于肝炎酶在手中，它将作为我们的初始 目标。然而，我们将追查与之密切相关的鼻病毒和 脊髓灰质炎病毒的酶通过同样的方法。我们预计每个系统 将为选择性抑制剂的设计提供新的见解。 人血清白蛋白溶酶体中的半胱氨酸蛋白酶 大多数人类细胞，组织蛋白酶B，H和L，将被用于研究 针对小核糖核酸病毒设计的化合物的特异性 蛋白酶。 将提供酶和第一代抑制剂的样品 给一个合作的结晶学小组。由此产生的数据 在进一步的努力中，将利用该方法来设计较新的抑制物 以提高选择性。

Our long term objective is to develop potent and selective inhibitors of the picornaviral processing enzymes that may be of value in antiviral therapy. To achieve this goal, we will study the catalytic and binding properties of the 3C cysteine proteinases of hepatitis A virus, human rhinovirus types 2 and 14 and human poliovirus. The collaborating group, led by Bruce Malcolm at Protos, has expressed large quantities of the hepatitis enzyme. We have examined a variety of potential peptide sub- strates and developed rapid, quantitative assays. We are now poised to expand our study by refining the peptide structure that is recognized by this enzyme, by developing new assay methods based on sensitive fluorescence methods, and by constructing potential inhibitors based on peptide structures. We will utilize all these methods to thoroughly analyze the interaction of substrates and inhibitors with the Hepatitis enzyme. As the hepatitis enzyme is in hand, it will serves as our initial target. However, we will pursue the closely related rhinovirus and poliovirus enzymes by the same approach. We anticipate that each system will provide new insights into selective inhibitors design. The human cysteine proteinases found in the lysosomal compartment of most human cells, cathepsins B,H, and L, will be utilized in studies for the specificity of compounds designed against the picornaviral proteinases. Samples of enzymes and first-generation inhibitor will be provided to a collaborating crystallography group. Data arising from this approach will be utilized to design newer inhibiotrs in a further effort to improve the selectivity.