Allosteric control of collagen fibril degradation by matrix metalloprotease-1

基质金属蛋白酶-1 对胶原原纤维降解的变构控制

• 批准号: 10853496
• 负责人: Susanta Kumar Sarkar
• 金额: $ 31.4万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10853496
• 关键词: Active Sites; Affect; Allosteric Site; Animals; Atherosclerosis; Binding; Biochemical; Biological Assay; Buffers; Catalysis; Catalytic Domain; Cause of Death; Cells; Collagen; Collagen Fibril; Collagen Type I; Communication; Complement; Disease; Distant; Drug usage; Dyes; Extracellular Matrix; Family; Fibrillar Collagen; Fluorescence Resonance Energy Transfer; Fluorescent Dyes; Gelatinase A; Goals; Health; Hemopexin; Home; Human; Human body; Image; Immobilization; Individual; Inflammation; Invaded; Kinetics; Label; Lead; Learning; Ligand Binding; Ligands; Location; MMP9 gene; Maps; Measures; Metalloproteases; Methods; Molecular Conformation; Morphology; Motion; Mutation; Neoplasm Metastasis; Organ; Pathologic; Pathologic Processes; Physiological; Play; Pneumonia; Proteins; Proteolysis; Public Health; Relaxation; Resistance; Resolution; Role; Scanning Electron Microscopy; Site; Specificity; Structure; Surface; Temperature; Testing; Tetracyclines; Therapeutic; Time; Tissues; Variant; Water; Weight; experimental study; fluorescence microscope; gel electrophoresis; improved; in vitro Assay; inhibitor; innovation; member; molecular dynamics; monomer; mutant; public health relevance; reconstitution; scaffold; screening; self assembly; side effect; simulation; single molecule; skin disorder; small molecule; therapeutic target; triple helix; wound

Project Summary Collagen, the most abundant human protein, provides a scaffold for cells to maintain tissue and organ integrity. Fibrillar collagen is highly resistant to proteolysis and is degraded by specific matrix metalloproteases (MMPs). The degradation of fibrillar collagen is an essential part of tissue remodeling and is involved in many normal and pathological processes. While the degradation of triple-helical collagen monomers is well-studied, degradation of native collagen fibrils remains poorly understood. Fibrils are insoluble in physiological buffers, heterogeneous, and extended substrates, making them challenging to study using standard biochemical assays. We have overcome these limitations by developing a single-molecule method to track and analyze the motion of labeled MMPs on fibrils using a home-built total internal reflection fluorescence microscope (TIRFM). We propose to study the roles of MMP1 and MMP9 in degrading type-1 collagen fibrils. MMP1 can degrade triple-helical collagen, whereas MMP9 cannot degrade triple-helical collagen significantly. However, MMP9 is upregulated in numerous skin diseases, cancer metastasis, wound, inflammation, pneumonia, and atherosclerosis. We propose to study the mechanism of fibril degradation by two important MMPs by a combination of single-molecule tracking, innovative analysis, simulations, ensemble assays, and animal studies.

项目摘要 胶原蛋白是人类最丰富的蛋白质，为细胞提供了一个支架，以维持组织和器官的完整性。 原纤维胶原对蛋白水解具有高度抗性，并被特定的基质金属蛋白酶（MMP）降解。 纤维胶原的降解是组织重塑的重要组成部分，并参与许多正常和 病理过程。虽然三螺旋胶原蛋白单体的降解已得到充分研究，但降解 对天然胶原纤维的结构仍知之甚少。原纤维不溶于生理缓冲液，是异质的， 和扩展的底物，使它们具有挑战性的研究使用标准的生物化学测定。我们有 通过开发单分子方法来跟踪和分析标记的运动， 使用自制的全内反射荧光显微镜（TIRFM）观察纤维上的MMP。我们建议 研究MMP 1和MMP 9在降解1型胶原纤维中的作用。MMP 1可降解三螺旋 胶原蛋白，而MMP 9不能显著降解三螺旋胶原。然而，MMP9是上调， 多种皮肤病、癌症转移、创伤、炎症、肺炎和动脉粥样硬化。我们提出 研究两种重要的MMPs通过单分子结合降解纤维的机制， 跟踪、创新分析、模拟、集成分析和动物研究。