Real-time measurements of kinetics of EGF binding to soluble EGF receptor monomers and dimers support the dimerization model for receptor activation

Zhou, Min; Felder, S; Rubinstein, Menachem; Hurwitz, David R.; Ullrich, Axel; Lax, Irit; Schlessinger, Joseph

doi:10.1021/bi00083a020

Cited by 128 publications

(91 citation statements)

References 23 publications

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“…Therefore, we conclude that the value of the off rate constant, k -1 , is unlikely to be much less than the value reported in Ref. 43. On the other hand, the on rate constant, k 1 , could be substantially higher for the receptor in situ because of the decreased orientation restrictions on the positions of encountering molecules.…”

Section: Derivation Of a Kinetic Modelcontrasting

confidence: 52%

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Quantification of Short Term Signaling by the Epidermal Growth Factor Receptor

Kholodenko¹,

Demin²,

Moehren³

et al. 1999

Journal of Biological Chemistry

532

605

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During the past decade, our knowledge of molecular mechanisms involved in growth factor signaling has proliferated almost explosively. However, the kinetics and control of information transfer through signaling networks remain poorly understood. This paper combines experimental kinetic analysis and computational modeling of the short term pattern of cellular responses to epidermal growth factor (EGF) in isolated hepatocytes. The experimental data show transient tyrosine phosphorylation of the EGF receptor (EGFR) and transient or sustained response patterns in multiple signaling proteins targeted by EGFR. Transient responses exhibit pronounced maxima, reached within 15-30 s of EGF stimulation and followed by a decline to relatively low (quasi-steady-state) levels. In contrast to earlier suggestions, we demonstrate that the experimentally observed transients can be accounted for without requiring receptor-mediated activation of specific tyrosine phosphatases, following EGF stimulation. The kinetic model predicts how the cellular response is controlled by the relative levels and activity states of signaling proteins and under what conditions activation patterns are transient or sustained. EGFR signaling patterns appear to be robust with respect to variations in many elemental rate constants within the range of experimentally measured values. On the other hand, we specify which changes in the kinetic scheme, rate constants, and total amounts of molecular factors involved are incompatible with the experimentally observed kinetics of signal transfer. Quantitation of signaling network responses to growth factors allows us to assess how cells process information controlling their growth and differentiation.

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Section: Derivation Of a Kinetic Modelcontrasting

confidence: 52%

“…, and the (43). The K d value of 0.6 nM, characteristic for membrane-bound receptor can be obtained if the reported (43) magnitude of k 1 is increased or k -1 is decreased by a factor of about 600.…”

Section: Derivation Of a Kinetic Modelmentioning

confidence: 99%

Quantification of Short Term Signaling by the Epidermal Growth Factor Receptor

Kholodenko¹,

Demin²,

Moehren³

et al. 1999

Journal of Biological Chemistry

532

605

View full text Add to dashboard Cite

show abstract

“…There the ligand is EGF, and the messenger is associated with calcium signaling and is able to pass through gap junctions; one possibility is inositol 1,4,5-trisphosphate (IP3). The cell size is a ∼ 10 μm, the dissociation constant is as small as (33), the dissociation rate is μ ∼ 0.001 s −1 (34), and the EGF diffusion constant is D ∼ 50 μm 2 s −1 (35). The turnover rate of IP3 is estimated as ν ∼ 0.1 s −1 (36).…”

Section: Discussionmentioning

confidence: 99%

Limits to the precision of gradient sensing with spatial communication and temporal integration

Mugler

Levchenko

Nemenman

2016

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

135

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Gradient sensing requires at least two measurements at different points in space. These measurements must then be communicated to a common location to be compared, which is unavoidably noisy. Although much is known about the limits of measurement precision by cells, the limits placed by the communication are not understood. Motivated by recent experiments, we derive the fundamental limits to the precision of gradient sensing in a multicellular system, accounting for communication and temporal integration. The gradient is estimated by comparing a "local" and a "global" molecular reporter of the external concentration, where the global reporter is exchanged between neighboring cells. Using the fluctuation-dissipation framework, we find, in contrast to the case when communication is ignored, that precision saturates with the number of cells independently of the measurement time duration, because communication establishes a maximum length scale over which sensory information can be reliably conveyed. Surprisingly, we also find that precision is improved if the local reporter is exchanged between cells as well, albeit more slowly than the global reporter. The reason is that whereas exchange of the local reporter weakens the comparison, it decreases the measurement noise. We term such a model "regional excitation-global inhibition." Our results demonstrate that fundamental sensing limits are necessarily sharpened when the need to communicate information is taken into account. C ells sense spatial gradients in environmental chemicals with remarkable precision. A single amoeba, for example, can respond to a difference of roughly 10 attractant molecules between the front and the back of the cell (1). Cells are even more sensitive when they are in a group: Cultures of many neurons respond to chemical gradients equivalent to a difference of only one molecule across an individual neuron's axonal growth cone (2), clusters of malignant lymphocytes have a wider chemotactic sensitivity than single cells (3), and groups of communicating epithelial cells detect gradients that are too weak for a single cell to detect (4). More generally, collective chemosensing properties are often very distinct from those in individual cells (3, 5-7). These observations have generated a renewed interest in the question of what sets the fundamental limit to the precision of gradient sensing in large, spatially extended, often collective sensory systems.Fundamentally, sensing a stationary gradient requires at least two measurements to be made at different points in space. The precision of these two or more individual measurements bounds the gradient sensing precision (8, 9). In its turn, each individual measurement is limited by the finite number of molecules within the detector volume and the ability of the detector to integrate over time, a point first made by Berg and Purcell (BP) (8). More detailed calculations of gradient sensing by specific geometries of receptors have since confirmed that the precision of gradient sensing remains limited by ...

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“…In addition to first-order kinetics, the model assumes that the concentrations of bound and unbound targeted tracer in the extravascular space are in instantaneous equilibrium. Although fundamental to the derivation of this model system, values of binding potential have been shown to be robust even when this assumption does not apply (42,43). Additional physiological processes, such as tracer metabolism and receptor internalization, are not captured by the model and therefore represent potential sources of error in the calculations of BP.…”

Section: Discussionmentioning

confidence: 99%

Dynamic dual-tracer MRI-guided fluorescence tomography to quantify receptor density in vivo

Davis

Samkoe²,

Tichauer

et al. 2013

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

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The up-regulation of cell surface receptors has become a central focus in personalized cancer treatment; however, because of the complex nature of contrast agent pharmacokinetics in tumor tissue, methods to quantify receptor binding in vivo remain elusive. Here, we present a dual-tracer optical technique for noninvasive estimation of specific receptor binding in cancer. A multispectral MRI-coupled fluorescence molecular tomography system was used to image the uptake kinetics of two fluorescent tracers injected simultaneously, one tracer targeted to the receptor of interest and the other tracer a nontargeted reference. These dynamic tracer data were then fit to a dual-tracer compartmental model to estimate the density of receptors available for binding in the tissue. Applying this approach to mice with deep-seated gliomas that overexpress the EGF receptor produced an estimate of available receptor density of 2.3 ± 0.5 nM (n = 5), consistent with values estimated in comparative invasive imaging and ex vivo studies.

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Real-time measurements of kinetics of EGF binding to soluble EGF receptor monomers and dimers support the dimerization model for receptor activation

Cited by 128 publications

References 23 publications

Quantification of Short Term Signaling by the Epidermal Growth Factor Receptor

Quantification of Short Term Signaling by the Epidermal Growth Factor Receptor

Limits to the precision of gradient sensing with spatial communication and temporal integration

Dynamic dual-tracer MRI-guided fluorescence tomography to quantify receptor density in vivo

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