Structure and Dynamics of Phase-Separated Proteins
Liquid-liquid phase separation (LLPS) of biomolecules has long been used to concentrate and encapsulate molecules and plays an important role in biology and biomaterials development. An increasing number of proteins have been found to undergo LLPS in response to environmental changes, and within cells can give rise to a variety of membraneless organelles that regulate a range of biochemical processes. A similar phenomenon has been observed in extracellular matrices, where LLPS of monomeric elastin leads to the deposition of cohesive layer droplets rich in hydrated proteins and cross-linking to form elastic matrices, an attractive platform for the development of responsive biomaterials with a wide range of applications. Despite the strong interest in proteins undergoing stimulus-responsive phase separation, high-resolution studies have been hampered by the highly disordered nature of proteins undergoing LLPS and the unfavorable kinetics caused by the increased viscosity in the phase-separated state.
Fig. 1. Interactions and regulatory mechanisms implicated in protein phase separation. (Boeynaems S, et al., 2018)
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The molecular structure and dynamics of the phase-separated state, the question of how and why these proteins form, and how their physical characteristics contribute to biological function have attracted our great interest. Our scientists are developing interdisciplinary approaches to determine the structure and dynamics of phase-separated proteins, accelerating your insight into their organization, molecular properties and regulation. We have leading technology platforms:
Nuclear Magnetic Resonance (NMR) Spectroscopy Platform
- Measures the transient formation of secondary structures, molecular motion and tumbling, and interactions of intrinsically disordered proteins.
- Characterizes the molecular motions on different time scales.
- Provides information on the single-strand nature of proteins undergoing LLPS and the interactions critical for assembly.
Solution-State NMR Spectroscopy Platform
- Provides detailed information on the structure and motion of individual components within biomolecular condensates.
X-Ray Diffraction Platform
Elastin-like polypeptides (ELPs) from protoelastin sequences also undergo LLPS, allowing the formation of biomaterials with material properties very similar to those of natural elastin. We use ELPs as a suitable proxy to study phase separation and elastin, providing X-ray diffraction methods to characterize the structural characterization of amyloid fibrils to analyze the LLPS behavior of elastin.
Scattering Methods Platform
We have scattering methods for different wavelengths of electromagnetic radiation to provide you with insight into the structure and organization of macromolecules in solution on length scales from tens of angstroms to microns.
Cryo-Electron Microscopy (cryo-EM) and Tomography (cryo-ET) Platform
- Provides structural information on length scales from Å to nm.
- Can be used for a wide variety of samples, including purified macromolecular complexes, cells, tissues or organisms.
- cryo-ET provides insight into the structural organization of mesoscale phase separated in vivo biomolecules.
CD BioSciences offers comprehensive structure and dynamics services for phase-separated proteins.
- Determination of Structure and Contacts for Mediating LLPS
- Analysis of Protein Motions and Conformational Changes in the Dispersed and Condensed Phases
Types of Samples to Study Protein Phase Separation
- Dispersed phase.
- Biphasic samples.
- Macroscopic condensed phase samples.
Our NMR data are widely used in LLPS studies of biomolecules. We offer differential labeling schemes for NMR spectroscopy of individual components to help our clients fully analyze the behavior and interactions of protein phases. Our methods can be used to characterize how small molecules interact with LLPS-preferring proteins, and to determine how they disrupt weak, multivalent interactions. If you are interested in our services, please do not hesitate to contact us for more information.
Reference
- Boeynaems S, et al. (2018) Protein Phase Separation: A New Phase in Cell Biology. Trends Cell Biol. 28(6):420-435.
For research use only, not intended for any clinical use.
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