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dc.contributor.authorPuma, Patricia
dc.contributor.authorBuxser, Stephen E.
dc.contributor.authorWatson, Leslie
dc.contributor.authorKelleher, Daniel J.
dc.contributor.authorJohnson, Gary L.
dc.date2022-08-11T08:08:00.000
dc.date.accessioned2022-08-23T15:39:00Z
dc.date.available2022-08-23T15:39:00Z
dc.date.issued1983-03-10
dc.date.submitted2014-10-22
dc.identifier.citationJ Biol Chem. 1983 Mar 10;258(5):3370-5.
dc.identifier.issn0021-9258 (Linking)
dc.identifier.pmid6298234
dc.identifier.urihttp://hdl.handle.net/20.500.14038/26058
dc.description.abstractThe receptor for nerve growth factor (NGF) has been purified to near homogeneity from octylglucoside extracts of A875 melanoma cell membranes by the use of repetitive affinity chromatography on NGF-Sepharose. Elution of purified receptor (NGF receptor) was accomplished with 0.15 M NaCl, pH 11.0, containing phosphatidylcholine and octylglucoside. Chromatography on two columns of NGF-Sepharose yielded a 1500-fold purification of the receptor, as assessed by 125I-NGF binding, and permitted recovery of 9% of the total binding activity in the soluble extract. Scatchard analysis of equilibrium binding of 125I-NGF provided similar Kd values for NGF receptors in soluble extracts of A875 membranes (2.2 nM) and with purified NGF receptor (3.1 nM). Examination of NGF receptor after electrophoresis on sodium dodecyl sulfate-polyacrylamide gels revealed the presence of two major peptides, of Mr = 85,000 and Mr = 200,000. Affinity labeling experiments, done with 125I-NGF and A875 cells, soluble extracts of A875 cell membranes, and purified receptor, show that both of these components of the NGF receptor can be specifically cross-linked to 125I-NGF.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=6298234&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://www.jbc.org/content/258/5/3370.full.pdf+html
dc.subjectCell Line
dc.subjectCell Membrane
dc.subjectChromatography, Affinity
dc.subjectHumans
dc.subjectKinetics
dc.subjectMelanoma
dc.subjectMolecular Weight
dc.subjectNerve Growth Factors
dc.subjectReceptors, Cell Surface
dc.subjectReceptors, Nerve Growth Factor
dc.subjectBiochemistry
dc.subjectBiochemistry, Biophysics, and Structural Biology
dc.subjectMolecular Biology
dc.titlePurification of the receptor for nerve growth factor from A875 melanoma cells by affinity chromatography
dc.typeJournal Article
dc.source.journaltitleThe Journal of biological chemistry
dc.source.volume258
dc.source.issue5
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/bmp_pp/198
dc.legacy.embargo2014-11-05T00:00:00-08:00
dc.identifier.contextkey6269472
html.description.abstract<p>The receptor for nerve growth factor (NGF) has been purified to near homogeneity from octylglucoside extracts of A875 melanoma cell membranes by the use of repetitive affinity chromatography on NGF-Sepharose. Elution of purified receptor (NGF receptor) was accomplished with 0.15 M NaCl, pH 11.0, containing phosphatidylcholine and octylglucoside. Chromatography on two columns of NGF-Sepharose yielded a 1500-fold purification of the receptor, as assessed by 125I-NGF binding, and permitted recovery of 9% of the total binding activity in the soluble extract. Scatchard analysis of equilibrium binding of 125I-NGF provided similar Kd values for NGF receptors in soluble extracts of A875 membranes (2.2 nM) and with purified NGF receptor (3.1 nM). Examination of NGF receptor after electrophoresis on sodium dodecyl sulfate-polyacrylamide gels revealed the presence of two major peptides, of Mr = 85,000 and Mr = 200,000. Affinity labeling experiments, done with 125I-NGF and A875 cells, soluble extracts of A875 cell membranes, and purified receptor, show that both of these components of the NGF receptor can be specifically cross-linked to 125I-NGF.</p>
dc.identifier.submissionpathbmp_pp/198
dc.contributor.departmentBiochemistry and Molecular Pharmacology
dc.contributor.departmentNeurobiology
dc.source.pages3370-5


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