Superagonistic Activation of ErbB-1 by EGF-related Growth Factors with Enhanced Association and Dissociation Rate Constants

Lenferink, Anne E.G.; Zoelen, E.J.J. van; Vugt, Marianne J.H. van; Grothé, Suzanne; Rotterdam, Walter van; Poll, Monique L.M. van de; O’Connor-McCourt, Maureen D.

doi:10.1016/s0021-9258(19)61439-0

Cited by 25 publications

(20 citation statements)

References 30 publications

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“…The IC50s for EGF stimulation using the final assay protocol (24-h incubation) were similar to values reported in the literature. 1,10,11 The EC50s for EGF with cells used 2 h after being plated out were lower than the EC50 for EGF for cells plated out 24 h before starting the assay. This might reflect cell growth over the 24-h incubation period.…”

Section: Discussionmentioning

confidence: 84%

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Evaluation of the InteraX™ System Technology in a High-Throughput Screening Environment

Büttner

Kumpf²,

Menzel³

et al. 2005

SLAS Discovery

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The authors have developed a cell-based high-throughput screening (HTS)-compatible assay to measure EGFR dimerization using the InteraX enzyme complementation technology of Applied Biosystems. The cells contain 2 chimeric proteins with complementing deletion mutants of the beta galactosidase enzyme, each fused to the extracellular and transmembrane part of EGFR. On binding of EGF, EGF receptor dimerizes and an active beta galactosidase is built. The authors used this homogeneous 384-well assay to screen about 20,000 diverse compounds. From 2 independent primary screen runs 239 hits were identified. For run 1, a mean S/B ratio of 4.26 and a mean Z' factor of 0.74 were obtained, for run 2 a mean S/B ratio of 3.88 and a mean Z' factor of 0.71 were obtained. After hit confirmation, repeated 4 times, 112 hits remained with a confirmation rate of 48.9%. Thirty of the 112 could be identified as cytotoxic. Fifty-one of the remaining 82 compounds could be shown to be inhibitors of the beta galactosidase enzyme itself. In summary, 31 compounds remained as potential EGFR dimerization or EGF stimulation inhibitors. The authors conclude that the InteraX system technology is HTS capable and can detect small molecule inhibitors capable of inhibiting protein-protein interactions.

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Section: Discussionmentioning

confidence: 84%

“…The results obtained were in the range reported in the literature. 1,10,11 The signal-tobackground (S/B) ratio was in the range of 5.0 to 6.4. Z′ factors were achieved between 0.69 and 0.89, with n = 4 for each concentration tested.…”

Section: Assay Development In 384-well Platesmentioning

confidence: 99%

Evaluation of the InteraX™ System Technology in a High-Throughput Screening Environment

Büttner

Kumpf²,

Menzel³

et al. 2005

SLAS Discovery

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“…In other words, EGFR-ligands with the fastest k off elicited the strongest biological response. 15,16 Therefore, we have used AFM to determine whether EGFR binding to crocidolite as a foreign "ligand" could elicit a k off response similar to that of published values for natural ligands produced in humans.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…In some instances, TGFα functions as a more potent agonist of EGFR. 16 The published k off values for EGFR binding to EGF range from 0.002 to 0.75 s −1 , and those for TGFα range from 0.04 to 2.3 s −1 (Table 3). [14][15][16]46 One series of k off values from Table 3 is particularly noteworthy.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Repetitive Dissociation from Crocidolite Asbestos Acts as Persistent Signal for Epidermal Growth Factor Receptor

et al. 2013

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Mesothelioma is an incurable form of cancer located most commonly in the pleural lining of the lungs and is associated almost exclusively with the inhalation of asbestos. The binding of asbestos to epidermal growth factor receptor (EGFR), a transmembrane signal protein, has been proposed as a trigger for downstream signaling of kinases and expression of genes involved in cell proliferation and inhibition of apoptosis. Here, we investigate the molecular binding of EGFR to crocidolite (blue asbestos; Na2(Fe(2+),Mg)3Fe2(3+)Si8O22(OH)2) in buffer solution. Atomic force microscopy measurements revealed an attractive force of interaction (i.e., bond) as EGFR was pulled from contact with long fibers of crocidolite. The rupture force of this bond increased with loading rate. According to the Bell model, the off-rate of bond dissociation (k(off)) for EGFR was 22 s(-1). Similar experiments with riebeckite crystals, the nonasbestiform variety of crocidolite, yielded a k(off) of 8 s(-1). These k(off) values on crocidolite and riebeckite are very rapid compared to published values for natural agonists of EGFR like transforming growth factor and epidermal growth factor. This suggests binding of EGFR to the surfaces of these minerals could elicit a response that is more potent than biological hormone or cytokine ligands. Signal transduction may cease for endogenous ligands due to endocytosis and subsequent degradation, and even riebeckite particles can be cleared from the lungs due to their short, equant habit. However, the fibrous habit of crocidolite leads to lifelong persistence in the lungs where aberrant, repetitious binding with EGFR may continually trigger the activation switch leading to chronic expression of genes involved in oncogenesis.

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“…2000)(45) described growth factor interactions with ErbB-1 in which the dynamic rate constants correlated better with mitogenic activity than did the equilibrium constants (42). A more pertinent example, however, is that of McDonnell et al(2001) (46), who identified a role for the Cε2 domain for IgE (see Figure2) in allergic responses.…”

mentioning

confidence: 99%

Approaches to Allergy Detection Using Aptasensors

Turner¹,

Minunni²,

Brys³

et al. 2006

Smart Biosensor Technology

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Superagonistic Activation of ErbB-1 by EGF-related Growth Factors with Enhanced Association and Dissociation Rate Constants

Cited by 25 publications

References 30 publications

Evaluation of the InteraX™ System Technology in a High-Throughput Screening Environment

Evaluation of the InteraX™ System Technology in a High-Throughput Screening Environment

Repetitive Dissociation from Crocidolite Asbestos Acts as Persistent Signal for Epidermal Growth Factor Receptor

Approaches to Allergy Detection Using Aptasensors

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