METALLOENZYMES AND CONNECTIVE TISSUE METABOLISM

金属酶和结缔组织代谢

• 批准号: 3275183
• 负责人: HAROLD E VAN WART
• 金额: $ 10.19万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-04-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3275183
• 关键词: acidity /alkalinity; affinity chromatography; aminoacid; carboxyl group; collagenase; connective tissue metabolism; enzyme structure; enzyme substrate; fluorimetry; metalloenzyme; 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 degradation of collagen under physiological conditions. Collagen, the major structural protein of connective tissue, is highly resistant to attack by all known digestive and intracellular enzymes. Hence, the action of collagenases is essential to all processes involving the degradation or remodeling of connective tissue. Collagenases, which are believed to be zinc metalloenzymes, will be isolated from various sources according to published procedures and by affinity chromatographic methods to be developed. 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 substitutions on enzymatic activity. The substrate specificities of these enzymes toward native collagen (Types I-IV) and towards a series of synthetic peptides will be examined in detail. Chemical modification 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.

这项研究的目的是探讨 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， 所有已知的消化酶和细胞内酶的攻击。 因此，行动 胶原酶是必不可少的所有过程涉及降解或 结缔组织重塑。 胶原酶，被认为是 锌金属酶，将从各种来源分离，根据 已发表的程序和亲和层析方法， 开发 所获得的酶将在以下方面表征： 它们的均匀性、金属含量、氨基酸组成、分子量 和亚基结构。 金属原子在催化中的作用将是 通过检查金属取代对酶的影响来研究 活动 这些酶对天然的底物特异性 胶原蛋白（I-IV型）和一系列合成肽将被 详细检查。 将进行化学改性反应， 鉴定必需氨基酸残基。 这项研究将促进 更好地理解胶原酶在哺乳动物结缔组织中的作用 组织代谢