TARGETS OF A MECHANISM-BASED PROBE AGAINST CYSTEINE PROTEASES

基于机制的半胱氨酸蛋白酶探针的靶标

• 批准号: 8169165
• 负责人: Tom Muir
• 金额: $ 0.12万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169165
• 关键词: Apoptosis; Apoptotic; Biochemical; Caspase; Cathepsins; Cathepsins B; Cell Death; Cells; Cessation of life; Chemicals; Computer Retrieval of Information on Scientific Projects Database; Cysteine Protease; Cytosol; Data; Development; Funding; Future; Grant; Homeostasis; Institution; Investigation; Knockout Mice; Lysosomes; Peptide Hydrolases; Publishing; Reading; Reporting; Research; Research Personnel; Resources; Role; Source; United States National Institutes of Health; Work; base; feeding; programs; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Using LC-MS/MS, we have identified the target of a mechanism-based probe against cysteine proteases upregulated during apopotosis as Cathepsin B, implicating its activity in cell death. Cells control their own death through a program termed apoptosis, which is indispensable for development and homeostasis in all metazoans. Lysosomal cysteine proteases are not normally thought of as participating in apoptosis; however, recent reports have shown that the cathepsin proteases can be released from the lysosome during apoptosis, where they can participate in cell death. We report here the development of an activity-based probe that, under optimized conditions, reports on cathepsin B activity only in apoptotic cells by reading out the release of cathepsin B from the lysosomes. Biochemical characterization of apoptosis in cells from cathepsin B null mice shows delayed and suboptimal activation of caspases. Our data further supports a role for cathepsin B in the cytosol as a positive regulator of a cell death feed-forward loop and provides a chemical tool for future investigations. This work has been published in Chem. Biol.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 使用LC-MS/MS，我们已经确定了基于机制的探针的靶标，该探针针对凋亡过程中上调的半胱氨酸蛋白酶作为组织蛋白酶B，这暗示了其在细胞死亡中的活性。 细胞通过称为凋亡的程序来控制自己的死亡，这对于所有后生动物中的发展和稳态都是必不可少的。通常认为溶酶体半胱氨酸蛋白酶参与凋亡。但是，最近的报道表明，在凋亡过程中可以从溶酶体中释放组织蛋白酶蛋白酶，在那里它们可以参与细胞死亡。我们在这里报告了基于活性的探针的开发，该探针在优化的条件下，仅通过读取从溶酶体中释放组织蛋白酶B的释放来报道组织蛋白酶B活性。来自组织蛋白酶b的细胞中细胞凋亡的生化表征显示caspase的延迟和次优地激活。我们的数据进一步支持了在细胞质中，组织蛋白酶B作为细胞死亡进料前环的积极调节剂的作用，并为将来的研究提供了化学工具。 这项工作已发表在化学中。生物。