INTERACTION OF ALCOHOL DEHYDROGENASE WITH F-ACTIN

本论文的工作旨在研究分子间的可逆相互作用， 乙醇脱氢酶与F-肌动蛋白，以及相关代谢产物的作用 他们的互动。 我们将测试是否催化性能的 结合的酶不同于游离酶， 结合酶具有不同的蛋白水解敏感性。 两匹马 将使用肝脏和酵母同工酶。 将通过以下方式研究相互作用： 使用超离心分离游离酶和结合酶， 上清液中游离酶的活性。 F-actin的作用 对乙醇脱氢酶周转率的影响 胰蛋白酶和糜蛋白酶消化酶的速度在 F-肌动蛋白的存在和不存在。 我们建议在一个更类似于 而不是缓冲溶液。 我们的工作将提供一个 对酒精的酶性质有更现实的理解 脱氢酶和酒精的氧化在肝脏和酵母细胞中， vivo. 我们的长期目标是研究改变 环境和细胞骨架成分对生物活性的影响 细胞质酶

The proposed work is designed to study the reversible interaction of alcohol dehydrogenase with F-actin, and the effect of relevant metabolites on their interaction. We will test whether the catalytic properties of the bound enzyme differ from those of the free enzyme, and whether the free and bound enzyme have different proteolytic susceptibility. Both the horse liver and yeast isozymes will be used. The interaction will be studied by separating the free and bound enzyme using Ultracentrifugation and assaying the activity of the free enzyme in the supernatant. The effect of F-actin on the turnover rate of alcohol dehydrogenase will be studied by comparing the rate of trypsin and chymotrypsin digestion of the enzyme in the presence and absence of F-actin. We are proposing to study alcohol dehydrogenase in a system more similar to a cellular environment than a buffer solution. Our work will provide a more realistic understanding of the enzymatic properties of alcohol dehydrogenase and the oxidation of alcohol in liver and yeast cells in vivo. Our long term objective is to study the effect of changing the environment and the cytoskeletal components on the biological activity of cytoplasmic enzymes.