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• W1866851443 endingPage "2313" @default.
• W1866851443 startingPage "2301" @default.
• W1866851443 abstract "Autocrine signaling is important in normal tissue physiology as well as pathological conditions. It is difficult to analyze these systems, however, because they are both self-contained and recursive. To understand how parameters such as ligand production and receptor expression influence autocrine activity, we investigated a human epidermal growth factor/epidermal growth factor receptor (EGF/EGFR) loop engineered into mouse B82 fibroblasts. We varied the level of ligand production using the tet-off expression system and used metalloprotease inhibitors to modulate ligand release. Receptor expression was varied using antagonistic blocking antibodies. We compared autocrine ligand release with receptor activation using a microphysiometer-based assay and analyzed our data using a quantitative model of ligand release and receptor dynamics. We found that the activity of our autocrine system could be described in terms of a simple ratio between the rate of ligand production (VLT) and the rate of receptor production (VR). At a VLT/VR ratio of &lt;0.3, essentially no ligand was found in the extracellular medium, but a significant number of cell receptors (30-40%) were occupied. As the VLT/VR ratio increased from 0.3 towards unity, receptor occupancy increased and significant amounts of ligand appeared in the medium. Above a VLT/VR ratio of 1.0, receptor occupancy approached saturation and most of the released ligand was lost into the medium. Analysis of human mammary epithelial cells showed that a VLT/VR ratio of &lt;5×10−4was sufficient to evoke &gt;20% of a maximal proliferative response. This demonstrates that natural autocrine systems can be active even when no ligand appears in the extracellular medium." @default.
• W1866851443 created "2016-06-24" @default.
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• W1866851443 date "2001-06-15" @default.
• W1866851443 modified "2023-10-03" @default.
• W1866851443 title "Quantitative analysis of the EGF receptor autocrine system reveals cryptic regulation of cell response by ligand capture" @default.
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• W1866851443 doi "https://doi.org/10.1242/jcs.114.12.2301" @default.
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