METALLOENZYMES AND COLLAGEN METABOLISM

金属酶和胶原蛋白代谢

• 批准号: 3071126
• 负责人: HAROLD E VAN WART
• 金额: $ 5.13万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-07-01 至 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3071126
• 关键词: affinity chromatography; collagenase; connective tissue metabolism; joint disorder; metalloenzyme; peptidases; rheumatoid arthritis; synovial fluid; zinc

The objective of the proposed research is to investigate the mechanism of action of collagenases, which are unique in their ability to catalyze the degradatio nof collagen under physiological conditions. Collagen, the major structural protein of connective tissue, is highly resistant to tattack by all known digestive and intracellular enzymes. Hence, the action of collagenase is essential to all processes involving the degradation or remodelling of connective tissues. In particular, they have been implicated in the enzymatic joint destructon that accompanies arthritis. Collagenases will be isolated from various sources according to published procedures and by affinity chromatographic methods to be developed. New and rapid fluorometric assay procedures will be developed to expedite the isolation and purification steps. The enzymes obtained will be characterized with respect to their homogeneity, metal content, amino acid composition, molecular weight and subunit structure. The role of the metal atom(s) in catalysis will be investigated by examining the effect of metal substituions on enzymatic activity. Chemical modificatin reactions will be carried out to identify essential amino acid residues. This research should promote a greater understanding of the role of collagenases in mammalian connective tissue metabolism.

拟议研究的目的是探讨 胶原酶的作用，这是独特的能力，催化 胶原蛋白在生理条件下的降解。 胶原 结缔组织的主要结构蛋白质，具有高度的抵抗力， 被所有已知的消化酶和细胞内酶攻击。 所以 胶原酶的作用对于所有涉及 结缔组织的退化或重塑。 特别是， 参与了伴随着 关节炎 胶原酶将根据以下方法从各种来源分离： 已发表的程序和亲和层析方法， 开发 将开发新的快速荧光测定程序 以加快分离和纯化步骤。 获得的酶 将以它们的均匀性，金属含量， 氨基酸组成、分子量和亚基结构。 的作用 金属原子在催化作用中的作用将通过检查 金属取代对酶活性的影响。 化学改性 进行反应以鉴定必需氨基酸残基。 这项研究应促进人们更好地了解 哺乳动物结缔组织代谢中的胶原酶。