Linked Life Data

7925442

Source:http://linkedlifedata.com/resource/pubmed/id/7925442

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1994-11-23
pubmed:abstractText
The binding of epidermal growth factor (EGF) or an EGF-like growth factor to the EGF receptor is the initial event which leads to receptor activation, and consequently the induction of cell growth. In order to study this binding interaction in detail, we produced the extracellular domain of the EGF receptor (EGFR) using the baculovirus expression system. Affinity-labeling and Western-blot analyses revealed that the baculovirus-infected insect cells secrete active EGFR extracellular domain relatively efficiently, however a significant amount of inactive EGFR extracellular domain is retained within the cells. The apparent dissociation constant (Kd) of the secreted EGFR extracellular domain for EGF and transforming growth factor alpha (TGF-alpha), as determined using an immobilized receptor binding assay, was approximately 200 nM. Interestingly, this Kd value is 30-40-fold lower than that of the full-length EGFR derived from detergent-solubilized A431 cell membranes. The stoichiometry of binding of the EGFR extracellular domain to EGF and TGF-alpha was examined by band-shift analysis on non-denaturing PAGE and was estimated to be 1:1. We have also shown, using sedimentation equilibrium analysis, that ligand binding induces significant dimerization of the EGFR extracellular domain. Finally, we carried out site-specific mutagenesis on the EGFR extracellular domain in order to define the ligand-binding region. We identified amino acid residues which are close to the binding site since they are common to the epitopes of several ligand-competitive monoclonal antibodies. However, these residues do not contribute directly to ligand binding since the affinity of the mutated EGFR extracellular domain for EGF and TGF-alpha was unaffected.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0014-2956
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
225
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
223-33
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:7925442-Amino Acid Sequence, pubmed-meshheading:7925442-Animals, pubmed-meshheading:7925442-Baculoviridae, pubmed-meshheading:7925442-Base Sequence, pubmed-meshheading:7925442-Binding Sites, pubmed-meshheading:7925442-Blotting, Western, pubmed-meshheading:7925442-Carcinoma, Squamous Cell, pubmed-meshheading:7925442-Cell Line, pubmed-meshheading:7925442-Chickens, pubmed-meshheading:7925442-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:7925442-Epidermal Growth Factor, pubmed-meshheading:7925442-Humans, pubmed-meshheading:7925442-Kinetics, pubmed-meshheading:7925442-Macromolecular Substances, pubmed-meshheading:7925442-Male, pubmed-meshheading:7925442-Mice, pubmed-meshheading:7925442-Molecular Sequence Data, pubmed-meshheading:7925442-Mutagenesis, Site-Directed, pubmed-meshheading:7925442-Receptor, Epidermal Growth Factor, pubmed-meshheading:7925442-Recombinant Proteins, pubmed-meshheading:7925442-Spodoptera, pubmed-meshheading:7925442-Transfection, pubmed-meshheading:7925442-Transforming Growth Factor alpha, pubmed-meshheading:7925442-Tumor Cells, Cultured
pubmed:year
1994
pubmed:articleTitle
The extracellular domain of the epidermal growth factor receptor. Studies on the affinity and stoichiometry of binding, receptor dimerization and a binding-domain mutant.
pubmed:affiliation
National Research Council of Canada, Biotechnology Research Institute, Montreal, Quebec.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't