scholarly journals Regulation of Transmembrane Signaling by Phase Separation

2019 ◽  
Vol 48 (1) ◽  
pp. 465-494 ◽  
Author(s):  
Lindsay B. Case ◽  
Jonathon A. Ditlev ◽  
Michael K. Rosen
Keyword(s):  
Phase Separation ◽  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Cluster Formation ◽  
Environmental Cues ◽  
Transmembrane Receptors ◽  
Domain Interactions ◽  
Micrometer Scale ◽  
Liquid Liquid Phase Separation ◽  
Oligomerization Domain

Cell surface transmembrane receptors often form nanometer- to micrometer-scale clusters to initiate signal transduction in response to environmental cues. Extracellular ligand oligomerization, domain-domain interactions, and binding to multivalent proteins all contribute to cluster formation. Here we review the current understanding of mechanisms driving cluster formation in a series of representative receptor systems: glycosylated receptors, immune receptors, cell adhesion receptors, Wnt receptors, and receptor tyrosine kinases. We suggest that these clusters share properties of systems that undergo liquid–liquid phase separation and could be investigated in this light.

scholarly journals Phase separation at the nanoscale quantified by dcFCCS

2020 ◽  
Vol 117 (44) ◽  
pp. 27124-27131
Author(s):  
Sijia Peng ◽  
Weiping Li ◽  
Yirong Yao ◽  
Wenjing Xing ◽  
Pilong Li ◽  
...  
Keyword(s):  
Phase Separation ◽  
Detection Limit ◽  
Fluorescence Microscopy ◽  
Critical Concentration ◽  
Transition Process ◽  
Correlation Spectroscopy ◽  
Cellular Processes ◽  
Conventional Fluorescence Microscopy ◽  
Micrometer Scale ◽  
Liquid Liquid Phase Separation

Liquid–liquid phase separation, driven by multivalent macromolecular interactions, causes formation of membraneless compartments, which are biomolecular condensates containing concentrated macromolecules. These condensates are essential in diverse cellular processes. Formation and dynamics of micrometer-scale phase-separated condensates are examined routinely. However, limited by commonly used methods which cannot capture small-sized free-diffusing condensates, the transition process from miscible individual molecules to micrometer-scale condensates is mostly unknown. Herein, with a dual-color fluorescence cross-correlation spectroscopy (dcFCCS) method, we captured formation of nanoscale condensates beyond the detection limit of conventional fluorescence microscopy. In addition, dcFCCS is able to quantify size and growth rate of condensates as well as molecular stoichiometry and binding affinity of client molecules within condensates. The critical concentration to form nanoscale condensates, identified by our experimental measurements and Monte Carlo simulations, is at least several fold lower than the detection limit of conventional fluorescence microscopy. Our results emphasize that, in addition to micrometer-scale condensates, nanoscale condensates are likely to play important roles in various cellular processes and dcFCCS is a simple and powerful quantitative tool to examine them in detail.

scholarly journals Specificity of LIM Domain Interactions with Receptor Tyrosine Kinases

1996 ◽  
Vol 271 (27) ◽  
pp. 15934-15941 ◽  
Author(s):  
Rui-yun Wu ◽  
Kyle Durick ◽  
Zhou Songyang ◽  
Lewis C. Cantley ◽  
Susan S. Taylor ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Lim Domain ◽  
Domain Interactions

scholarly journals ROR and RYK extracellular region structures suggest that receptor tyrosine kinases have distinct WNT-recognition modes

2021 ◽  
Author(s):  
Fumin Shi ◽  
Jeannine M Mendrola ◽  
Joshua B Sheetz ◽  
Neo Wu ◽  
Anselm Sommer ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Acyl Chain ◽  
Lipid Binding ◽  
Receptor Interaction ◽  
Transmembrane Receptors ◽  
Binding Characteristics ◽  
Interaction Sites ◽  
Rich Domain ◽  
Extracellular Region

WNTs play key roles in development and disease, by binding both Frizzled (FZD) seven-pass transmembrane receptors and numerous co-receptors that include the ROR and RYK receptor tyrosine kinases (RTKs). We describe crystal structures and WNT-binding characteristics of extracellular regions from the Drosophila ROR and RYK orthologs Nrk (neurospecific receptor tyrosine kinase) and Derailed-2 (Drl-2). RORs bind WNTs though a FZD-related cysteine-rich domain (CRD), and RYKs through a WNT-inhibitory factor (WIF) domain. Our structures suggest that neither the Nrk CRD nor the Drl-2 WIF domain can accommodate the acyl chain typically attached to WNTs. The Nrk CRD contains a deeply buried bound fatty acid, unlikely to be exchangeable with a WNT acyl chain. The Drl-2 WIF domain lacks the lipid-binding site seen in WIF-1. We also show that DWnt-5, which regulates Drosophila ROR and RYK orthologs, lacks an acyl chain. Together with analysis of WNT/receptor interaction sites, these structures provide new insight into how WNTs recruit their RTK co-receptors into signaling complexes.

scholarly journals Structural Aspects of Transmembrane Domain Interactions of Receptor Tyrosine Kinases

2011 ◽  
Vol 100 (3) ◽  
pp. 207a ◽  
Author(s):  
Eduard Bocharov ◽  
Konstantin Mineev ◽  
Dmitry Lesovoy ◽  
Marina Goncharuk ◽  
Sergey Goncharuk ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Transmembrane Domain ◽  
Domain Interactions ◽  
Structural Aspects

scholarly journals Solubility product constant directs the formation of biomolecular condensates

2020 ◽  
Author(s):  
Aniruddha Chattaraj ◽  
Michael L. Blinov ◽  
Leslie M. Loew
Keyword(s):  
Phase Separation ◽  
Solubility Product ◽  
Cluster Formation ◽  
Cellular Structures ◽  
Stochastic Network ◽  
Physical Chemical ◽  
Solubility Product Constant ◽  
Concentration Threshold ◽  
Large Clusters ◽  
Liquid Liquid Phase Separation

AbstractBiomolecular condensates, formed by liquid-liquid phase separation (LLPS), are important cellular structures. Using stochastic network-free kinetic models, we establish a physical-chemical basis for the concentration threshold of heterotypic multivalent molecules required for LLPS. We associate phase separation with a bimodal partitioning of the cluster distribution into small oligomers vs. huge polymers. The simulations reveal that LLPS obeys the solubility product constant (Ksp): the product of monomer concentrations, accounting for ideal stoichiometries, does not exceed a threshold no matter how much additional monomer is added to the system – additional monomer is funneled into large clusters. The Ksp applies over a range of valencies and stoichiometries. However, consistent with the importance of disordered domains for LLPS, removing flexible linker domains funnels valency-matched monomers into a “dimer trap”, and Ksp no longer defines a threshold for large cluster formation. We propose Ksp as a new tool for elucidating biomolecular condensate biophysics.

The role of cluster formation and metastable liquid—liquid phase separation in protein crystallization

2012 ◽  
Vol 159 ◽  
pp. 313 ◽  
Author(s):  
Fajun Zhang ◽  
Felix Roosen-Runge ◽  
Andrea Sauter ◽  
Roland Roth ◽  
Maximilian W. A. Skoda ◽  
...  
Keyword(s):  
Phase Separation ◽  
Liquid Phase ◽  
Protein Crystallization ◽  
Cluster Formation ◽  
Liquid Phase Separation ◽  
Metastable Liquid ◽  
Liquid Liquid Phase Separation

scholarly journals Synergistic Phase Separation of Two Pathways Promotes Integrin Clustering and Integrin Adhesion Complex Formation

2021 ◽  
Author(s):  
Michael K Rosen ◽  
Lindsay B. Case ◽  
Lisa Henry
Keyword(s):  
Phase Separation ◽  
Focal Adhesion ◽  
Environmental Cues ◽  
Physical Mechanisms ◽  
Environmental Perturbations ◽  
Integrin Clustering ◽  
Adhesion Complex ◽  
Liquid Liquid Phase Separation ◽  
In Cells

Integrin adhesion complexes (IACs) are integrin-based plasma membrane-associated comp iartments where cells sense environmental cues. The physical mechanisms and molecular interactions that mediate nascent IAC formation are unclear. We found that both p130Cas ("Cas") and Focal adhesion kinase ("FAK") undergo liquid-liquid phase separation in vitro under physiologic conditions. Cas- and FAK- driven phase separation is sufficient to reconstitute kindlindependent integrin clustering in vitro. In vitro condensates and cellular IACs exhibit similar sensitivities to environmental perturbations including changes in temperature and pH. Furthermore, mutations that inhibit or enhance phase separation in vitro reduce or increase the number of IACs in cells, respectively. Finally, we find that the Cas and FAK pathways act synergistically to promote phase separation, integrin clustering and IAC formation in vitro and in cells. We propose that Cas- and FAK- driven phase separation provides an intracellular trigger for integrin clustering and nascent IAC formation.

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