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Elucidating mechanisms of amyloid nucleation in vivo

阐明体内淀粉样成核机制

基本信息

批准号：
9886371
负责人：
Randal Arthur Halfmann
金额：
$ 31.76万
依托单位：
STOWERS INSTITUTE FOR MEDICAL RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9886371
关键词：
Adsorption Age Aging Alzheimer&apos s Disease Amyloid Biochemical Biophysics Cells Cellular biology Cytology Data Dependence Disease Event Flow Cytometry Fluorescence Resonance Energy Transfer Genetic Polymorphism Goals Grant Investigation Kinetics Laboratories Lead Longevity Memory Molecular Molecular Conformation Mutation Nature Nerve Degeneration Neurodegenerative Disorders Organism Outcome Pathologic Pathologic Processes Phase Physical condensation Physiological Physiological Processes Polymers Polymorph Population Probability Proteins Research Role Signal Transduction Solubility Specificity Structure Testing Therapeutic Thermodynamics Time amyloid formation amyloid structure analytical tool density environmental change experimental study fluorophore in vivo insight novel therapeutics polypeptide protein function proteostasis screening stress granule tool

项目摘要

Project Summary Amyloids exert numerous physiological and pathological activities. On the one hand, they perpetuate molecular memories and transduce intracellular signals; on the other, they precipitate incurable neurodegenerative and age-associated diseases. These disparate activities unfold over time scales that exceed the lifespan of the proteins or even the cells that harbor them. Our long-term goal is to determine whether, and how, these time scales emerge from the kinetics of amyloid formation in living cells. Toward this goal, we have focused on the critical first step of amyloid formation: nucleation. The probabilistic nature of nucleation has made its study exceedingly difficult with established cytological tools. We therefore developed Distributed Amphifluoric FRET (DAmFRET) to quantify nucleation as a function of a given protein’s concentration in living cells. We have now used DAmFRET to analyze nucleation of diverse amyloids and related polymers, and these data lead us to propose the central hypothesis of this grant: low-specificity interactions critically influence the rates and structural outcomes of amyloid nucleation in vivo. This hypothesis makes the following predictions that will be tested in our Specific Aims: that nucleation will favor increasingly labile amyloids with 1) increasing protein concentration or 2) condensation of the protein; and 3) that the ability of amyloids of one protein to nucleate amyloids of a different protein depends on their disordered content rather than their structure. Completing this investigation will reveal critical mechanistic features of nucleation by diverse proteins in living cells, including the thermodynamic underpinnings of nucleation barriers, and networks of pathological cross-seeding. It will also have opened a conduit to fundamental physical insights that are presently beyond the reach of cell biology. These will yield a deeper understanding of, and ultimately new therapeutic options for, age- associated and neurodegenerative diseases.

项目总结