Liquid–Liquid Phase Separation Modifies the Dynamic Properties of Intrinsically Disordered Proteins

• Published in: Journal of the American Chemical Society Vol. 145; no. 19; pp. 10548 - 10563
• Main Authors: Guseva, Serafima, Schnapka, Vincent, Adamski, Wiktor, Maurin, Damien, Ruigrok, Rob W. H., Salvi, Nicola, Blackledge, Martin
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 17.05.2023
• Subjects: Intrinsically Disordered Proteins - chemistry; Magnetic Resonance Spectroscopy; Molecular Dynamics Simulation; Motion; Protein Conformation
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/jacs.2c13647

Liquid–liquid phase separation of flexible biomolecules has been identified as a ubiquitous phenomenon underlying the formation of membraneless organelles that harbor a multitude of essential cellular processes. We use nuclear magnetic resonance (NMR) spectroscopy to compare the dynamic properties of an intrinsically disordered protein (measles virus N TAIL) in the dilute and dense phases at atomic resolution. By measuring 15N NMR relaxation at different magnetic field strengths, we are able to characterize the dynamics of the protein in dilute and crowded conditions and to compare the amplitude and timescale of the different motional modes to those present in the membraneless organelle. Although the local backbone conformational sampling appears to be largely retained, dynamics occurring on all detectable timescales, including librational, backbone dihedral angle dynamics and segmental, chainlike motions, are considerably slowed down. Their relative amplitudes are also drastically modified, with slower, chain-like motions dominating the dynamic profile. In order to provide additional mechanistic insight, we performed extensive molecular dynamics simulations of the protein under self-crowding conditions at concentrations comparable to those found in the dense liquid phase. Simulation broadly reproduces the impact of formation of the condensed phase on both the free energy landscape and the kinetic interconversion between states. In particular, the experimentally observed reduction in the amplitude of the fastest component of backbone dynamics correlates with higher levels of intermolecular contacts or entanglement observed in simulations, reducing the conformational space available to this mode under strongly self-crowding conditions.