Size-Based Chromatography of Signaling Clusters in a Living Cell Membrane

Caculitan, Niña G.; Kai, Hiroyuki; Liu, Eulanca Y.; Fay, Nicole C.; Ye, Yu; Lohmüller, Theobald; O’Donoghue, Geoff P.; Groves, Jay T.

doi:10.1021/nl404514e

Cited by 21 publications

(21 citation statements)

References 41 publications

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“…Finally, the adhesion and growth of primary neuronal cells cultured on these positively charged SLBs on gold is comparable with that of conventional protein coatings. Our presented procedure could be used for the formation of biomimetic surfaces on micro/ nanofabricated gold electrodes that can be used to measure electrical and optical signals of cellular interaction in biologically 61 and physically [62][63][64] diverse systems.…”

Section: Discussionmentioning

confidence: 99%

Positively charged supported lipid bilayer formation on gold surfaces for neuronal cell culture

et al. 2016

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Supported lipid bilayers are widely used as cell membrane models and sensor platforms, but the usage on gold surface needs additional surface modification or optimized experimental conditions. In this work, the authors show lipid bilayer formation on plasma activated gold surfaces in physiological conditions without any other modification if at least 30% positively charged lipids are present. Details of bilayer formation from small unilamellar vesicles were monitored using quartz crystal microbalance with dissipation in both basic and acidic environment. The authors also confirmed that this positively charged bilayer system can sustain primary cortical neuron growth and lipid transfer. This method will provide simple means to construct biomimetic interface on gold electrodes.

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Section: Discussionmentioning

confidence: 99%

Positively charged supported lipid bilayer formation on gold surfaces for neuronal cell culture

et al. 2016

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“…Supported lipid bilayers are suited for such studies because they allow membrane-associated proteins to be organized into micron-scale assemblies, a process that can be physically perturbed by the application of nanopatterned substrates to elucidate the role of assembly in signal transduction (24)(25)(26)(27)(28). Mobility-dependent assembly, and physical perturbation of such assemblies could not be achieved together in other cell adhesion assays such as monolayer cell cultures or those that involve display of protein on solid surfaces.…”

Section: Introductionmentioning

confidence: 99%

Sustained α -catenin Activation at E-cadherin Junctions in the Absence of Mechanical Force

Biswas

Hartman

Zaidel-Bar

et al. 2016

Biophysical Journal

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Mechanotransduction at E-cadherin junctions has been postulated to be mediated in part by a force-dependent conformational activation of a-catenin. Activation of a-catenin allows it to interact with vinculin in addition to F-actin, resulting in a strengthening of junctions. Here, using E-cadherin adhesions reconstituted on synthetic, nanopatterned membranes, we show that activation of a-catenin is dependent on E-cadherin clustering, and is sustained in the absence of mechanical force or association with F-actin or vinculin. Adhesions were formed by filopodia-mediated nucleation and micron-scale assembly of E-cadherin clusters, which could be distinguished as either peripheral or central assemblies depending on their relative location at the cell-bilayer adhesion. Whereas F-actin, vinculin, and phosphorylated myosin light chain associated only with the peripheral assemblies, activated a-catenin was present in both peripheral and central assemblies, and persisted in the central assemblies in the absence of actomyosin tension. Impeding filopodia-mediated nucleation and micron-scale assembly of E-cadherin adhesion complexes by confining the movement of bilayer-bound E-cadherin on nanopatterned substrates reduced the levels of activated a-catenin. Taken together, these results indicate that although the initial activation of a-catenin requires micron-scale clustering that may allow the development of mechanical forces, sustained force is not required for maintaining a-catenin in the active state.

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“…Some examples include the growth factor receptors (1,2), components pertaining to actin cytoskeleton reorganization (3), Ephrin receptors (4)(5)(6)(7)(8)(9), and membrane receptors in T cells (10)(11)(12). In each of these systems, the signaling molecules contain repetitive multivalent protein-protein interaction domains that drive the extended assembly of molecular complexes during signal transduction.…”

mentioning

confidence: 99%

Phosphotyrosine-mediated LAT assembly on membranes drives kinetic bifurcation in recruitment dynamics of the Ras activator SOS

Huang

Yan

Lin

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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118

174

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The assembly of cell surface receptors with downstream signaling molecules is a commonly occurring theme in multiple signaling systems. However, little is known about how these assemblies modulate reaction kinetics and the ultimate propagation of signals. Here, we reconstitute phosphotyrosine-mediated assembly of extended linker for the activation of T cells (LAT):growth factor receptor-bound protein 2 (Grb2):Son of Sevenless (SOS) networks, derived from the T-cell receptor signaling system, on supported membranes. Single-molecule dwell time distributions reveal two, well-differentiated kinetic species for both Grb2 and SOS on the LAT assemblies. The majority fraction of membrane-recruited Grb2 and SOS both exhibit fast kinetics and single exponential dwell time distributions, with average dwell times of hundreds of milliseconds. The minor fraction exhibits much slower kinetics, extending the dwell times to tens of seconds. Considering this result in the context of the multistep process by which the Ras GEF (guanine nucleotide exchange factor) activity of SOS is activated indicates that kinetic stabilization from the LAT assembly may be important. This kinetic proofreading effect would additionally serve as a stochastic noise filter by reducing the relative probability of spontaneous SOS activation in the absence of receptor triggering. The generality of receptor-mediated assembly suggests that such effects may play a role in multiple receptor proximal signaling processes.signal transduction | protein assembly | kinetic proofreading | membrane dwell time | single molecule

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Size-Based Chromatography of Signaling Clusters in a Living Cell Membrane

Cited by 21 publications

References 41 publications

Positively charged supported lipid bilayer formation on gold surfaces for neuronal cell culture

Positively charged supported lipid bilayer formation on gold surfaces for neuronal cell culture

Sustained α -catenin Activation at E-cadherin Junctions in the Absence of Mechanical Force

Phosphotyrosine-mediated LAT assembly on membranes drives kinetic bifurcation in recruitment dynamics of the Ras activator SOS

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