37,579 research outputs found
Impact of receptor clustering on ligand binding
<p>Abstract</p> <p>Background</p> <p>Cellular response to changes in the concentration of different chemical species in the extracellular medium is induced by ligand binding to dedicated transmembrane receptors. Receptor density, distribution, and clustering may be key spatial features that influence effective and proper physical and biochemical cellular responses to many regulatory signals. Classical equations describing this kind of binding kinetics assume the distributions of interacting species to be homogeneous, neglecting by doing so the impact of clustering. As there is experimental evidence that receptors tend to group in clusters inside membrane domains, we investigated the effects of receptor clustering on cellular receptor ligand binding.</p> <p>Results</p> <p>We implemented a model of receptor binding using a Monte-Carlo algorithm to simulate ligand diffusion and binding. In some simple cases, analytic solutions for binding equilibrium of ligand on clusters of receptors are provided, and supported by simulation results. Our simulations show that the so-called "apparent" affinity of the ligand for the receptor decreases with clustering although the microscopic affinity remains constant.</p> <p>Conclusions</p> <p>Changing membrane receptors clustering could be a simple mechanism that allows cells to change and adapt its affinity/sensitivity toward a given stimulus.</p
The impact of high density receptor clusters on VEGF signaling
Vascular endothelial growth factor (VEGF) signaling is involved in the
process of blood vessel development and maintenance. Signaling is initiated by
binding of the bivalent VEGF ligand to the membrane-bound receptors (VEGFR),
which in turn stimulates receptor dimerization. Herein, we discuss experimental
evidence that VEGF receptors localize in caveloae and other regions of the
plasma membrane, and for other receptors, it has been shown that receptor
clustering has an impact on dimerization and thus also on signaling. Overall,
receptor clustering is part of a complex ecosystem of interactions and how
receptor clustering impacts dimerization is not well understood. To address
these questions, we have formulated the simplest possible model. We have
postulated the existence of a single high affinity region in the cell membrane,
which acts as a transient trap for receptors. We have defined an ODE model by
introducing high- and low-density receptor variables and introduce the
corresponding reactions from a realistic model of VEGF signal initiation.
Finally, we use the model to investigate the relation between the degree of
VEGFR concentration, ligand availability, and signaling. In conclusion, our
simulation results provide a deeper understanding of the role of receptor
clustering in cell signaling.Comment: In Proceedings HSB 2013, arXiv:1308.572
Effects of receptor clustering on ligand dissociation: Theory and simulations
Receptor-ligand binding is a critical first step in signal transduction and
the duration of the interaction can impact signal generation. In mammalian
cells, clustering of receptors may be facilitated by heterogeneous zones of
lipids, known as lipid rafts. In vitro experiments show that disruption of
rafts significantly alters the dissociation of fibroblast growth factor-2
(FGF-2) from heparan sulfate proteoglycans, co-receptors for FGF-2. In this
paper, we develop a continuum stochastic formalism in order to (i) study how
rebinding affects the dissociation of ligands from a planar substrate, and (ii)
address the question of how receptor clustering influences ligand rebinding. We
find that clusters reduce the effective dissociation rate dramatically when the
clusters are dense and the overall surface density of receptors is low. The
effect is much less pronounced in the case of high receptor density and shows
non-monotonic behavior with time. These predictions are verified via lattice
Monte Carlo simulations. Comparison with experimental results suggests that the
theory does not capture the complete biological system. We speculate that
additional co-operative mechanisms might be present in order to increase ligand
retention, and present one possible ``internal diffusion'' model.Comment: Expanded text and added figures, revised version to appear in
Biophys.
GibbsCluster: unsupervised clustering and alignment of peptide sequences
Receptor interactions with short linear peptide fragments (ligands) are at the base of many biological signaling processes. Conserved and information-rich amino acid patterns, commonly called sequence motifs, shape and regulate these interactions. Because of the properties of a receptor-ligand system or of the assay used to interrogate it, experimental data often contain multiple sequence motifs. GibbsCluster is a powerful tool for unsupervised motif discovery because it can simultaneously cluster and align peptide data. The GibbsCluster 2.0 presented here is an improved version incorporating insertion and deletions accounting for variations in motif length in the peptide input. In basic terms, the program takes as input a set of peptide sequences and clusters them into meaningful groups. It returns the optimal number of clusters it identified, together with the sequence alignment and sequence motif characterizing each cluster. Several parameters are available to customize cluster analysis, including adjustable penalties for small clusters and overlapping groups and a trash cluster to remove outliers. As an example application, we used the server to deconvolute multiple specificities in large-scale peptidome data generated by mass spectrometry.Fil: Andreatta, Massimo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); ArgentinaFil: Alvarez, Bruno. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); ArgentinaFil: Nielsen, Morten. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Technical University of Denmark; Dinamarc
Integrin activation - the importance of a positive feedback
Integrins mediate cell adhesion and are essential receptors for the
development and functioning of multicellular organisms. Integrin activation is
known to require both ligand and talin binding and to correlate with cluster
formation but the activation mechanism and precise roles of these processes are
not yet resolved. Here mathematical modeling, with known experimental
parameters, is used to show that the binding of a stabilizing factor, such as
talin, is alone insufficient to enable ligand-dependent integrin activation for
all observed conditions; an additional positive feedback is required.Comment: in press in Bulletin of Mathematical Biolog
Sampling of conformational ensemble for virtual screening using molecular dynamics simulations and normal mode analysis
Aim: Molecular dynamics simulations and normal mode analysis are
well-established approaches to generate receptor conformational ensembles
(RCEs) for ligand docking and virtual screening. Here, we report new fast
molecular dynamics-based and normal mode analysis-based protocols combined with
conformational pocket classifications to efficiently generate RCEs. Materials
\& methods: We assessed our protocols on two well-characterized protein targets
showing local active site flexibility, dihydrofolate reductase and large
collective movements, CDK2. The performance of the RCEs was validated by
distinguishing known ligands of dihydrofolate reductase and CDK2 among a
dataset of diverse chemical decoys. Results \& discussion: Our results show
that different simulation protocols can be efficient for generation of RCEs
depending on different kind of protein flexibility
A role for Syk-kinase in the control of the binding cycle of the β2 integrins (CD11/CD18) in human polymorphonuclear neutrophils
A fine control of β2 integrin (CD11/CD18)-mediated firm adhesion of human neutrophils to the endothelial cell monolayer is required to allow ordered emigration. To elucidate the molecular mechanisms that control this process, intracellular protein tyrosine signaling subsequent to β2 integrin-mediated ligand binding was studied by immunoprecipitation and Western blotting techniques. The 72-kDa Syk-kinase, which was tyrosine-phosphorylated upon adhesion, was found to coprecipitate with CD18, the β-subunit of the β2 integrins. Moreover, inhibition of Syk-kinase by piceatannol enhanced adhesion and spreading but diminished N-formyl-Met-Leu-Phe-induced chemotactic migration. The enhancement of adhesiveness was associated with integrin clustering, which results in increased integrin avidity. In contrast, piceatannol had no effect on the surface expression or on the affinity of β2 integrins. Altogether, this suggests that Syk-kinase controls alternation of β2 integrin-mediated ligand binding with integrin detachment
Self-consistent theory of reversible ligand binding to a spherical cell
In this article, we study the kinetics of reversible ligand binding to
receptors on a spherical cell surface using a self-consistent stochastic
theory. Binding, dissociation, diffusion and rebinding of ligands are
incorporated into the theory in a systematic manner. We derive explicitly the
time evolution of the ligand-bound receptor fraction p(t) in various regimes .
Contrary to the commonly accepted view, we find that the well-known
Berg-Purcell scaling for the association rate is modified as a function of
time. Specifically, the effective on-rate changes non-monotonically as a
function of time and equals the intrinsic rate at very early as well as late
times, while being approximately equal to the Berg-Purcell value at
intermediate times. The effective dissociation rate, as it appears in the
binding curve or measured in a dissociation experiment, is strongly modified by
rebinding events and assumes the Berg-Purcell value except at very late times,
where the decay is algebraic and not exponential. In equilibrium, the ligand
concentration everywhere in the solution is the same and equals its spatial
mean, thus ensuring that there is no depletion in the vicinity of the cell.
Implications of our results for binding experiments and numerical simulations
of ligand-receptor systems are also discussed.Comment: 23 pages with 4 figure
Mechanotransduction and growth factor signalling to engineer cellular microenvironments
Engineering cellular microenvironments involves biochemical factors, the extracellular matrix (ECM) and the interaction with neighbouring cells. This progress report provides a critical overview of key studies that incorporate growth factor (GF) signalling and mechanotransduction into the design of advanced microenvironments. Materials systems have been developed for surface-bound presentation of GFs, either covalently tethered or sequestered through physico-chemical affinity to the matrix, as an alternative to soluble GFs. Furthermore, some materials contain both GF and integrin binding regions and thereby enable synergistic signalling between the two. Mechanotransduction refers to the ability of the cells to sense physical properties of the ECM and to transduce them into biochemical signals. Various aspects of the physics of the ECM, i.e. stiffness, geometry and ligand spacing, as well as time-dependent properties, such as matrix stiffening, degradability, viscoelasticity, surface mobility as well as spatial patterns and gradients of physical cues are discussed. To conclude, various examples illustrate the potential for cooperative signalling of growth factors and the physical properties of the microenvironment for potential applications in regenerative medicine, cancer research and drug testing
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