SynDrops · Understanding the physiological and pathological relevance of liquid-liquid phase separation by synuclein family of proteins

Understanding the physiological and pathological relevance of liquid-liquid phase separation by synuclein family of proteins

Context and Overall Objectives of the Project: Project SynDrops investigates the phase separation behavior of α-synuclein (α-Syn), a protein associated with Parkinson's disease (PD) and other neurodegenerative disorders. The primary focus is on how pathological mutations and terminal truncations affect the thermodynamics and physical properties of α-Syn phase condensates and their role in modulating amyloid aggregation. Key Background and Motivation: Understanding the phase separation behavior of α-Syn, particularly how mutations and truncations influence this process, is crucial as it is believed to be a precursor to amyloid fibril formation—a hallmark of synucleinopathies. In addition, recent findings of nanoscale phase separation challenge classical theories, suggesting the presence of stable, small molecular clusters below the critical concentration for protein phase separation. We also discover and characterize these nanoscale assemblies for α-Syn and study the effects of pathological mutations on the nanoscale phase separation landscape. Advanced Techniques Utilized: 1. Capflex and TDIPS: Techniques developed on the FIDA1 instrument to measure phase separation properties. 2. Mass Photometry: Employed to detect and characterize nanoscale phase separation of α-Syn. Primary Objectives: 1. Impact of Pathological Mutations: Determine how familial mutations in α-Syn influence its phase separation properties, focusing on dilute phase concentrations and associated free energy changes (ΔG). 2. Examine Terminal Truncations: Investigate the effects of truncating the C-terminal region of α-Syn on its phase behavior and stability. 3. Characterize Nanocluster Formation: Explore the formation of α-Syn micelles and nanoclusters at varying ionic strengths and their transition into larger condensates. 4. Material Properties of Condensates: Analyze the material properties of α-Syn condensates, such as viscosity and resistance to dissolution. 5. Quantify Amyloid Fibril Formation: Develop methods to estimate amyloid fibril concentration within α-Syn condensates and correlate this with phase separation dynamics. Project Pathway to Impact: Scientific and Technical Impact: • Understanding Disease Mechanisms: By elucidating how mutations and truncations influence α-Syn phase separation and aggregation, SynDrops aims to uncover the mechanisms driving neurodegenerative diseases. • Novel Techniques: The validation of Capflex, TDIPS and mass photometry techniques provides valuable tools for broader applications in cellular and molecular biology in the context of phase separation. Expected Contributions: 1. Enhanced Scientific Knowledge: Advancing our understanding of protein phase behavior and its implications for disease. 2. Therapeutic Development: Informing the design of interventions that can modulate protein phase behavior, offering new avenues for combating neurodegenerative disorders. 3. Diagnostic Tools: Detailed characterization of α-Syn nanoclusters and condensates could lead to new strategies for diagnosing and treating synucleinopathies by targeting early-stage aggregation events.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз