CARBOHYDRATE PROTEIN CONJUGATES OF PROTEASES

蛋白酶的碳水化合物蛋白缀合物

• 批准号: 3307182
• 负责人: JOHN S SWENTON
• 金额: $ 16.83万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1994-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3307182
• 关键词: carbohydrates; chemical conjugate; chemical structure; chemical synthesis; endopeptidases; peptide chemical synthesis

In this proposal, we outline the synthesis of new materials that stabilize proteases in organic solvents for their use in the catalytic formation of peptide bonds. We detail the synthesis of these new carbohydrate based materials, the preparation of carbohydrate protein conjugates of proteases [CPC(proteases)], and explore the use of these materials for the enzyme catalyzed synthesis of oligo- and polypeptides in organic solvent systems. Our investigations on the use of CPC (proteases) should extend to other enzymes and may lead to new opportunities for the use of proteins in organic synthesis. Specifically, our goals include: * The synthesis of a series of carbohydrate-functionalized macromolecules that contain masked aldehyde functionality, with charged, hydrophobic or hydrophilic groups. We propose the preparation of carbohydrate-based macromolecules that contain functionality that can effectively control the surface properties of proteases to preserve their catalytic activity in organic solvents. * The synthesis and characterization of carbohydrate protein conjugates of proteases. We plan to study conjugates of proteases for two reasons: (1) proteases are well-characterized enzymes (X-ray crystallographic data, mechanistic detail, etc.) which may allow us to develop a fundamental understanding of the mechanism of stabilization of these enzymes conjugates in organic solvents, and (2) proteases are important for the development of convergent. methodology in polypeptide synthesis. * The development of carbohydrate protein conjugates of proteases [CPC(proteases)] for convergent polypeptide synthesis. We will study the fundamental problems associated with the enzymatic synthesis of peptide bonds: hydrolysis of activated substrates, transamidation, substrate specificity, etc. We will also use CPC(proteases) for the coupling of large peptide fragments in organic solvents (and organic solvent-water mixtures) such as acetonitrile, dimethylformamide and dimethylsulfoxide with the overall goal being the development of the convergent synthesis of polypeptides and proteins.

在该提案中，我们概述了新材料的合成， 在有机溶剂中稳定蛋白酶，用于催化 形成肽键。 我们详细介绍了这些新的合成 碳水化合物基材料，碳水化合物蛋白的制备 蛋白酶的缀合物[CPC（蛋白酶）]，并探索这些缀合物的用途。 用于酶催化合成寡肽和多肽的材料 在有机溶剂体系中。 我们对CPC（蛋白酶）使用的研究应该扩展到其他领域。 酶，并可能导致新的机会，利用蛋白质在 有机合成具体而言，我们的目标包括： * 一系列碳水化合物功能化的 含有掩蔽的醛官能团的大分子，带有电荷， 疏水或亲水基团。 我们建议编制 基于碳水化合物的大分子，其含有可 有效地控制蛋白酶的表面性质， 在有机溶剂中的催化活性。 * 糖蛋白的合成与表征 蛋白酶的缀合物。 我们计划研究蛋白酶的结合物， 两个原因：（1）蛋白酶是表征良好的酶（X射线 晶体学数据、机械细节等）这也许能让我们 从根本上了解稳定的机制， 这些酶在有机溶剂中的缀合物，和（2）蛋白酶， 这对融合的发展很重要。多肽方法学 合成. * 蛋白酶糖蛋白偶联物的研究进展 [CPC（蛋白酶）]用于会聚多肽合成。 我们会研究 与肽的酶促合成相关的基本问题 键：活化底物的水解，转酰胺作用，底物 我们还将使用CPC（蛋白酶）用于偶联 在有机溶剂（和有机溶剂-水）中的大肽片段 混合物）如乙腈、二甲基甲酰胺和二甲亚砜 总的目标是发展聚合合成 多肽和蛋白质。