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Large-scale fluid/fluid phase separation of proteins and lipids in giant plasma membrane vesicles

TLDR
It is demonstrated that giant plasma membrane vesicles (GPMVs) or blebs formed from the plasma membranes of cultured mammalian cells can also segregate into micrometer-scale fluid phase domains, and GPMVs now provide an effective approach to characterize biological membrane heterogeneities.
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Crosslinking a lipid raft component triggers liquid ordered-liquid disordered phase separation in model plasma membranes.

TLDR
This work investigates the large-scale physical effect of crosslinking a minor membrane component, the ganglioside GM1, in simple lipid models of the plasma membrane containing sphingomyelin, cholesterol, and phosphatidylcholine, and finds that this lipid separation causes a dramatic redistribution of a transmembrane peptide, consistent with a raft model of membrane organization.
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A Lipid Raft Environment Enhances Lyn Kinase Activity by Protecting the Active Site Tyrosine from Dephosphorylation*

TLDR
It is proposed that lipid rafts exclude a phosphatase that negatively regulates Lyn kinase activity by constitutive dephosphorylation of the kinase domain tyrosine residue of Lyn in a lipid raft environment.
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Critical fluctuations in plasma membrane vesicles.

TLDR
Critical behavior in giant plasma membrane vesicles that are isolated directly from living cells suggest that the compositions of mammalian plasma membranes are tuned to reside near a miscibility critical point and that heterogeneity corresponding to < 50 nm-sized compositional fluctuations are present in GPMV membranes at physiological temperatures.
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Cancer cell-derived microvesicles induce transformation by transferring tissue transglutaminase and fibronectin to recipient cells

TLDR
It is demonstrated that MV shed by two different human cancer cells are capable of conferring onto normal fibroblasts and epithelial cells the transformed characteristics of cancer cells and that this effect requires the transfer of the protein cross-linking enzyme tissue transglutaminase (tTG).
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Correlation Functions Quantify Super-Resolution Images and Estimate Apparent Clustering Due to Over-Counting

TLDR
An analytical method using correlation functions to quantify clustering in super-resolution fluorescence localization images and electron microscopy images of static surfaces in two dimensions is presented and it is demonstrated that over-counting does not give rise to apparent co-clustering in double label experiments when pair cross-correlation functions are measured.
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Compartmentalized Activation of the High Affinity Immunoglobulin E Receptor within Membrane Domains*

TLDR
It is reported here that aggregation-dependent association of FcεRI with specialized regions of the plasma membrane precedes its tyrosine phosphorylation and appears necessary for this event, and participation of membrane domains accounts for previously unexplained aspects of F cεRI-mediated signaling.
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Bimolecular Fluorescence Complementation ( BiFC ) Analysis as a Probe of Protein Interactions in Living Cells

TLDR
Bimolecular fluorescence complementation analysis enables direct visualization of protein interactions in living cells and can be performed using standard molecular biology and cell culture reagents and a regular fluorescence microscope or flow cytometer.

Structural determinants for partitioning of lipids and proteins between coexisting fluid phases in giant plasma membrane vesicles.

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Fluorescence anisotropy measurements of lipid order in plasma membranes and lipid rafts from RBL-2H3 mast cells.

TLDR
It is shown using model membranes with varying amounts of cholesterol that steady-state fluorescence anisotropy is a sensitive measure of cholesterol-dependent ordering, and the results suggest that DPH-PC is a more sensitive probe than NBD-PE.
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