SMALL MOLECULE CATALYST OF PROTEIN FOLDING

蛋白质折叠的小分子催化剂

• 批准号: 6298203
• 负责人: RONALD T RAINES
• 金额: $ 0.75万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6298203
• 关键词: bioengineering /biomedical engineering; biomedical resource; microorganism; nuclear magnetic resonance spectroscopy

Protein disulfide isomerase (PDI) catalyzed the interchange of disulfide bonds in substrate proteins. The two Cys-Gly-His-Cys active sites provide a thiol of low pKa and a disulfide bond of high reduction potential (E??). Trans-1,20bis(2-mercaptoacetamido)cyclohexane (BMC) mimics the pKa and E?? of the PDI active sites. In vitro assays demonstrate that BMC is an efficient catalyst of the reactivation of scrambled ribonuclease (Rnase) A, a substrate with non-native disulfide bonds. In vivo assays show that BMC in the growth medium of yeast cells increases the secretion of native acid phosphatase, which has eight disulfide bonds. The effect is similar to that from the overproduction of PDI in the yeast cells. A monothiol analog of BMC, N-methyl-mercaptoacetamide (NMA), also catalyzes the reactivation of scrambled Rnase A, but less efficiently than does BMC. These data indicate that molecules with a low thiol pKa and high disulfide E?? catalyze effectively the formation of native disulfide bonds in proteins. Moreover, BMC is a useful catalyst for the in vitro and in vivo folding of proteins that contain multiple disulfide bonds.

蛋白质二硫键异构酶（PDI）催化的交换 底物蛋白中的二硫键。 两个 Cys-Gly-His-Cys 活性 位点提供低 pKa 的硫醇和高 pKa 的二硫键 还原电位（E??）。 Trans-1,20bis(2-巯基乙酰氨基)环己烷 (BMC) 模拟 pKa 和 乙?? PDI 活性位点。 体外试验表明 BMC 是 重新激活乱序核糖核酸酶的有效催化剂 (Rnase) A，具有非天然二硫键的底物。 体内 分析表明，酵母细胞生长培养基中的 BMC 可以提高 分泌天然酸性磷酸酶，具有八个二硫键。 其效果与 PDI 生产过剩的效果类似。 酵母细胞。 BMC 的单硫醇类似物，N-甲硫基乙酰胺 (NMA)，也催化乱序RNA酶A的重新激活，但作用较少 比 BMC 更高效。 这些数据表明具有 低硫醇 pKa 和高二硫键 E??有效催化 蛋白质中天然二硫键的形成。 此外，BMC 是 蛋白质体外和体内折叠的有用催化剂 含有多个二硫键。