Comparing the Epidermal Growth Factor Interaction with Four Different Cell Lines: Intriguing Effects Imply Strong Dependency of Cellular Context

Björkelund, Hanna; Gedda, Lars; Andersson, Karl

doi:10.1371/journal.pone.0016536

Cited by 48 publications

(60 citation statements)

References 21 publications

(24 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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.

136

View full text Add to dashboard Cite

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 ...

show abstract

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.

136

View full text Add to dashboard Cite

show abstract

“…We have previously shown that the kinetic behavior of the EGF-EGFR interaction may vary greatly among cell lines (14), as demonstrated by real-time interaction measurements performed using LigandTracer ® instruments. The binding curves showed at least two parallel interaction events, one fast-on-fast-off interaction and one higher affinity interaction, in which the association and dissociation rates of EGF were much slower.…”

Section: Introductionmentioning

confidence: 94%

“…Previous studies have demonstrated that the presence of gefitinib affects the kinetic properties (association and dissociation rates) of the EGF-EGFR interaction in certain cell lines, observed as an increase in affinity (10,14). This observation may be the result of the larger number of EGFR dimers present on the cell surface upon gefitinib treatment, regardless of their kinase activity.…”

Section: Introductionmentioning

confidence: 98%

Resolving the EGF-EGFR interaction characteristics through a multiple-temperature, multiple-inhibitor, real-time interaction analysis approach

Björkelund

Gedda

Malmqvist³

et al. 2012

Molecular and Clinical Oncology

Self Cite

View full text Add to dashboard Cite

Overexpression and aberrant activity of the epidermal growth factor (EGF) have been observed in various cancer types, rendering it an important target in oncology research. The interaction between EGF and its receptor (EGFR), as well as subsequent internalization, is complex and may be affected by various factors including tyrosine kinase inhibitors (TKIs). By combining real-time binding curves produced in LigandTracer with internalization assays conducted at different temperatures and with different TKIs, the processes of ligand binding, internalization and excretion was visualized. SKOV3 cells had a slower excretion rate compared to A431 and U343 cells, and the tested TKIs (gefitinib, lapatinib, AG1478 and erlotinib) reduced the degree of internalization. The kinetic analysis of the binding curves further demonstrated TKI-dependent balances of EGFR monomer and dimer populations, where lapatinib promoted the monomeric form, while the other TKIs induced dimers. The dimer levels were found to be associated with the apparent affinity of the EGF-EGFR interaction, with EGF binding stronger to EGFR dimers compared to monomers. This study analyzed how real-time molecular interaction analysis may be utilized in combination with perturbations in order to understand the kinetics of a ligand-receptor interaction, as well as some of its associated intracellular processes. Our multiple-temperature and -inhibitor assay setup renders it possible to follow the EGFR monomer, dimer and internalized populations in a detailed manner, allowing for a new perspective of the EGFR biology.

show abstract

“…LigandTracer Yellow was used for 131 I-cetuximab and Affibody molecule 111 In-Z342 measurements, and LigandTracer Grey for 125 I-trastuzumab measurements, all according to previously published protocol (3). After a short baseline measurement, the cells were incubated first at a lower concentration of protein, then at a higher concentration and finally the cells were put in fresh cell culture medium to monitor the release of bound material.…”

Section: Real-time Interaction Measurements In Ligandtracermentioning

confidence: 99%

Protein interactions with HER-family receptors can have different characteristics depending on the hosting cell line

Andersson¹

2011

Int J Oncol

Self Cite

View full text Add to dashboard Cite

Abstract.Cell lines are common model systems in the development of therapeutic proteins and in the research on cellular functions and dysfunctions. In this field, the protein interaction assay is a frequently used tool for assessing the adequacy of a protein for diagnostic and therapeutic purposes. In this study, we investigated the extent to which the interaction characteristics depend on the choice of cell line for HER-family receptors. The interaction characteristics of two therapeutic antibodies (trastuzumab and cetuximab) and one Affibody molecule (Z HER2:342 ), interacting with the intended receptor were characterized with high precision using an automated realtime interaction method, in different cell lines (HaCaT, A431, HEP-G2, SKOV3, PC3, DU-145). Clear differences in binding affinity and kinetics, up to one order of magnitude, were found for the interaction of the same protein binding to the same receptor on different cells for all three proteins. For HER-family receptors, it is therefore important to refer to the measured affinity for a protein-receptor interaction together with the hosting cell line. The ability to accurately measure affinity and kinetics of a protein-receptor interaction on cell lines of different origins may increase the understanding of underlying receptor biology, and impact the selection of candidates in the development of therapeutic or diagnostic agents.

show abstract

Comparing the Epidermal Growth Factor Interaction with Four Different Cell Lines: Intriguing Effects Imply Strong Dependency of Cellular Context

Cited by 48 publications

References 21 publications

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

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

Resolving the EGF-EGFR interaction characteristics through a multiple-temperature, multiple-inhibitor, real-time interaction analysis approach

Protein interactions with HER-family receptors can have different characteristics depending on the hosting cell line

Contact Info

Product

Resources

About