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dc.contributor.authorBuxser, Stephen E.
dc.contributor.authorKelleher, Daniel J.
dc.contributor.authorWatson, Leslie
dc.contributor.authorPuma, Patricia
dc.contributor.authorJohnson, Gary L.
dc.date2022-08-11T08:08:00.000
dc.date.accessioned2022-08-23T15:38:59Z
dc.date.available2022-08-23T15:38:59Z
dc.date.issued1983-03-25
dc.date.submitted2014-10-22
dc.identifier.citationJ Biol Chem. 1983 Mar 25;258(6):3741-9.
dc.identifier.issn0021-9258 (Linking)
dc.identifier.pmid6300053
dc.identifier.urihttp://hdl.handle.net/20.500.14038/26057
dc.description.abstractThe binding of 125I-labeled nerve growth factor (NGF) to human melanoma cell (A875) membranes, detergent-soluble membrane extracts, and membrane extracts reconstituted into phospholipid vesicles was significantly increased when binding was carried out in the presence of wheat germ agglutinin (WGA). In the absence of WGA, all 125I-NGF binding was rapidly eliminated by trypsin treatment or rapidly dissociated in the presence of a high concentration of unlabeled NGF. However, in the presence of WGA, up to 75% of 125I-NGF bound was resistant to trypsin digestion and was only slowly dissociated by a high concentration of unlabeled NGF. The effects of WGA can be blocked or reversed by N-acetylglucosamine. Both WGA and NGF rapidly associate with soluble extracts and reconstituted vesicles and, at the concentrations used here, reach binding equilibrium within 2 min. The conversion to slowly dissociating, trypsin-resistant binding, however, was not complete for at least 10 min. Both WGA and NGF are required for maximum accumulation of trypsin-resistant, slowly dissociating binding. The order of addition of NGF and WGA has no effect on the rate of conversion of NGF-receptor, and the conversion occurs after both NGF and WGA are present. The amount of conversion is dependent on the incubation temperature, and significantly greater conversion occurs at 37 than at 0 degrees C. The generation of the trypsin-resistant, slowly dissociating state of NGF-receptor is consistent with a time- and temperature-dependent conformational change in NGF-receptor which occurs after interaction of both NGF and WGA with the receptor or closely associated structures.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=6300053&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://www.jbc.org/content/258/6/3741.long
dc.subjectCell Line
dc.subjectCell Membrane
dc.subjectHumans
dc.subjectKinetics
dc.subjectLectins
dc.subjectLiposomes
dc.subjectMelanoma
dc.subjectNerve Growth Factors
dc.subjectReceptors, Cell Surface
dc.subjectReceptors, Nerve Growth Factor
dc.subjectTrypsin
dc.subjectWheat Germ Agglutinins
dc.subjectBiochemistry
dc.subjectBiochemistry, Biophysics, and Structural Biology
dc.subjectMolecular Biology
dc.titleChange in state of nerve growth factor receptor. Modulation of receptor affinity by wheat germ agglutinin
dc.typeJournal Article
dc.source.journaltitleThe Journal of biological chemistry
dc.source.volume258
dc.source.issue6
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/bmp_pp/197
dc.legacy.embargo2014-11-05T00:00:00-08:00
dc.identifier.contextkey6269471
html.description.abstract<p>The binding of 125I-labeled nerve growth factor (NGF) to human melanoma cell (A875) membranes, detergent-soluble membrane extracts, and membrane extracts reconstituted into phospholipid vesicles was significantly increased when binding was carried out in the presence of wheat germ agglutinin (WGA). In the absence of WGA, all 125I-NGF binding was rapidly eliminated by trypsin treatment or rapidly dissociated in the presence of a high concentration of unlabeled NGF. However, in the presence of WGA, up to 75% of 125I-NGF bound was resistant to trypsin digestion and was only slowly dissociated by a high concentration of unlabeled NGF. The effects of WGA can be blocked or reversed by N-acetylglucosamine. Both WGA and NGF rapidly associate with soluble extracts and reconstituted vesicles and, at the concentrations used here, reach binding equilibrium within 2 min. The conversion to slowly dissociating, trypsin-resistant binding, however, was not complete for at least 10 min. Both WGA and NGF are required for maximum accumulation of trypsin-resistant, slowly dissociating binding. The order of addition of NGF and WGA has no effect on the rate of conversion of NGF-receptor, and the conversion occurs after both NGF and WGA are present. The amount of conversion is dependent on the incubation temperature, and significantly greater conversion occurs at 37 than at 0 degrees C. The generation of the trypsin-resistant, slowly dissociating state of NGF-receptor is consistent with a time- and temperature-dependent conformational change in NGF-receptor which occurs after interaction of both NGF and WGA with the receptor or closely associated structures.</p>
dc.identifier.submissionpathbmp_pp/197
dc.contributor.departmentBiochemistry and Molecular Pharmacology
dc.contributor.departmentNeurobiology
dc.source.pages3741-9


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