A simple luciferase assay for signal transduction activity detection of epidermal growth factor displayed on phage.

Studies on receptor-ligand interactions are important for the design of agonists or antagonists of natural ligands. We developed a luciferase reporter assay to screen epidermal growth factor receptor (EGFR) binding molecules rapidly for their ability to stimulate or inhibit signal transduction. Huma...

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Main Authors: Souriau, C, Fort, P, Roux, P, Hartley, O, Lefranc, M P, Weill, M
Formato: Artigo
Idioma:en
Publicado em: 1997
Assuntos:
Research Article
Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC146613/
https://ncbi.nlm.nih.gov/pubmed/9092666
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spelling pubmed-1466132003-04-01 A simple luciferase assay for signal transduction activity detection of epidermal growth factor displayed on phage. Souriau, C Fort, P Roux, P Hartley, O Lefranc, M P Weill, M Nucleic Acids Res Research Article Studies on receptor-ligand interactions are important for the design of agonists or antagonists of natural ligands. We developed a luciferase reporter assay to screen epidermal growth factor receptor (EGFR) binding molecules rapidly for their ability to stimulate or inhibit signal transduction. Human EGF displayed on fd filamentous phage presented an activity similar to soluble EGF when tested for binding to the EGFR, for induction of cell cycle progression or in the luciferase assay. Two libraries of human EGF variants displayed on phage were constructed in which the aspartic acid residue at position 46 or the arginine residue at position 41 were randomised. EGF mutants displayed on phage were screened in parallel for binding to the EGFR using an ELISA assay and for transducing activity using the luciferase assay. Regarding the 46 position, most of the mutants retained the ability to bind the EGFR and their transducing activity corresponded perfectly with their binding. For the more crucial 41 position, only the wild-type EGF was able to bind the EGFR. Our approach allowed a simple determination of crucial positions and paved the way for identification of agonists with altered transduction activity. 1997-04-15 /pmc/articles/PMC146613/ /pubmed/9092666 Text en
institution US National Library of Medicine
collection PubMed Central
language en
format Article
topic Research Article
spellingShingle Research Article
Souriau, C
Fort, P
Roux, P
Hartley, O
Lefranc, M P
Weill, M
A simple luciferase assay for signal transduction activity detection of epidermal growth factor displayed on phage.
description Studies on receptor-ligand interactions are important for the design of agonists or antagonists of natural ligands. We developed a luciferase reporter assay to screen epidermal growth factor receptor (EGFR) binding molecules rapidly for their ability to stimulate or inhibit signal transduction. Human EGF displayed on fd filamentous phage presented an activity similar to soluble EGF when tested for binding to the EGFR, for induction of cell cycle progression or in the luciferase assay. Two libraries of human EGF variants displayed on phage were constructed in which the aspartic acid residue at position 46 or the arginine residue at position 41 were randomised. EGF mutants displayed on phage were screened in parallel for binding to the EGFR using an ELISA assay and for transducing activity using the luciferase assay. Regarding the 46 position, most of the mutants retained the ability to bind the EGFR and their transducing activity corresponded perfectly with their binding. For the more crucial 41 position, only the wild-type EGF was able to bind the EGFR. Our approach allowed a simple determination of crucial positions and paved the way for identification of agonists with altered transduction activity.
author Souriau, C
Fort, P
Roux, P
Hartley, O
Lefranc, M P
Weill, M
author_facet Souriau, C
Fort, P
Roux, P
Hartley, O
Lefranc, M P
Weill, M
author_sort Souriau, C
title A simple luciferase assay for signal transduction activity detection of epidermal growth factor displayed on phage.
title_short A simple luciferase assay for signal transduction activity detection of epidermal growth factor displayed on phage.
title_full A simple luciferase assay for signal transduction activity detection of epidermal growth factor displayed on phage.
title_fullStr A simple luciferase assay for signal transduction activity detection of epidermal growth factor displayed on phage.
title_full_unstemmed A simple luciferase assay for signal transduction activity detection of epidermal growth factor displayed on phage.
title_sort simple luciferase assay for signal transduction activity detection of epidermal growth factor displayed on phage.
publishDate 1997
url https://ncbi.nlm.nih.gov/pmc/articles/PMC146613/
https://ncbi.nlm.nih.gov/pubmed/9092666
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