Immunoaffinity purification of membrane protein constituents of the IM-9 lymphoblast receptor for substance P
Substance P (SP) is an undecapeptide neuromediator that stimulates human T-lymphocyte function by binding to stereospecific membrane receptors. Human IM-9 cultured B-lymphoblasts express approximately 20,000 receptors per cell for [ 125I]SP with a K d of 0.3 nM. [ 125I]SP was specifically crosslinke...
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| Veröffentlicht in: | Analytical biochemistry 1987-08, Vol.164 (2), p.502-513 |
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| container_title | Analytical biochemistry |
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| creator | McGillis, Joseph P. Organist, Michele L. Payan, Donald G. |
| description | Substance P (SP) is an undecapeptide neuromediator that stimulates human T-lymphocyte function by binding to stereospecific membrane receptors. Human IM-9 cultured B-lymphoblasts express approximately 20,000 receptors per cell for [
125I]SP with a
K
d
of 0.3 nM. [
125I]SP was specifically crosslinked by disuccinimidyl suberate to IM-9 cell membrane proteins of 78, 58, and 33 kDa. An indirect immunoaffinity purification procedure has now been developed based on immunoabsorption of detergent-solubilized [
125I]SP-labeled IM-9 cell membrane proteins to anti-SP antibody that was bound to an epoxide ultraffinity high-performance liquid chromatography column, followed by elution in acidic 8
m urea. The 58- and 33-kDa SP-receptor complexes were purified to apparent homogeneity by immunoaffinity chromatography and identified by autoradiography and silver staining of sodium dodecyl sulfate-polyacrylamide gels. |
| doi_str_mv | 10.1016/0003-2697(87)90525-2 |
| format | Article |
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125I]SP with a
K
d
of 0.3 nM. [
125I]SP was specifically crosslinked by disuccinimidyl suberate to IM-9 cell membrane proteins of 78, 58, and 33 kDa. An indirect immunoaffinity purification procedure has now been developed based on immunoabsorption of detergent-solubilized [
125I]SP-labeled IM-9 cell membrane proteins to anti-SP antibody that was bound to an epoxide ultraffinity high-performance liquid chromatography column, followed by elution in acidic 8
m urea. The 58- and 33-kDa SP-receptor complexes were purified to apparent homogeneity by immunoaffinity chromatography and identified by autoradiography and silver staining of sodium dodecyl sulfate-polyacrylamide gels.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/0003-2697(87)90525-2</identifier><identifier>PMID: 2823637</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Affinity Labels ; Cell Line ; Chromatography, Affinity - methods ; Chromatography, High Pressure Liquid - methods ; Humans ; immunoaffinity purification ; lymphocyte ; Lymphocytes - analysis ; lymphocytes T ; man ; Membrane Proteins - isolation & purification ; Molecular Weight ; Receptors, Neurokinin-1 ; Receptors, Neurotransmitter - isolation & purification ; Solubility ; substance P ; substance P receptor</subject><ispartof>Analytical biochemistry, 1987-08, Vol.164 (2), p.502-513</ispartof><rights>1987</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c388t-5f7f5d15a492a0ee03309a51c0aa3d21bd85b035eca9a4b63ea355fa0bd89b803</citedby><cites>FETCH-LOGICAL-c388t-5f7f5d15a492a0ee03309a51c0aa3d21bd85b035eca9a4b63ea355fa0bd89b803</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0003-2697(87)90525-2$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2823637$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>McGillis, Joseph P.</creatorcontrib><creatorcontrib>Organist, Michele L.</creatorcontrib><creatorcontrib>Payan, Donald G.</creatorcontrib><title>Immunoaffinity purification of membrane protein constituents of the IM-9 lymphoblast receptor for substance P</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>Substance P (SP) is an undecapeptide neuromediator that stimulates human T-lymphocyte function by binding to stereospecific membrane receptors. Human IM-9 cultured B-lymphoblasts express approximately 20,000 receptors per cell for [
125I]SP with a
K
d
of 0.3 nM. [
125I]SP was specifically crosslinked by disuccinimidyl suberate to IM-9 cell membrane proteins of 78, 58, and 33 kDa. An indirect immunoaffinity purification procedure has now been developed based on immunoabsorption of detergent-solubilized [
125I]SP-labeled IM-9 cell membrane proteins to anti-SP antibody that was bound to an epoxide ultraffinity high-performance liquid chromatography column, followed by elution in acidic 8
m urea. The 58- and 33-kDa SP-receptor complexes were purified to apparent homogeneity by immunoaffinity chromatography and identified by autoradiography and silver staining of sodium dodecyl sulfate-polyacrylamide gels.</description><subject>Affinity Labels</subject><subject>Cell Line</subject><subject>Chromatography, Affinity - methods</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>Humans</subject><subject>immunoaffinity purification</subject><subject>lymphocyte</subject><subject>Lymphocytes - analysis</subject><subject>lymphocytes T</subject><subject>man</subject><subject>Membrane Proteins - isolation & purification</subject><subject>Molecular Weight</subject><subject>Receptors, Neurokinin-1</subject><subject>Receptors, Neurotransmitter - isolation & purification</subject><subject>Solubility</subject><subject>substance P</subject><subject>substance P receptor</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1987</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU2LFDEQhoMo6-zoP1DISfTQWkk63Z2LIIuuAyt60HNIpytspNNpk7Qw_96MM-xRD0Ud3re-niLkBYO3DFj3DgBEwzvVvx76Nwoklw1_RHYMVNeAAPWY7B4sT8l1zj8BGGtld0Wu-MBFJ_odCYcQtiUa5_ziy5GuW_LOW1N8XGh0NGAYk1mQrikW9Au1ccnFlw2Xkk-Gco_08KVRdD6G9T6Os8mFJrS4lpioq5G3MRezWKTfnpEnzswZn1_ynvz49PH7zefm7uvt4ebDXWPFMJRGut7JiUnTKm4AEUQ9x0hmwRgxcTZOgxxBSLRGmXbsBBohpTNQBTUOIPbk1blv3frXhrno4LPFea6XxC3roUJSlc1_jawdBFcV1Z60Z6NNMeeETq_JB5OOmoE-vUOfWOsTaz30-u87NK9lLy_9tzHg9FB04V_192cdK43fHpPO1mNlNfnKsOgp-n8P-AMiQ5s9</recordid><startdate>19870801</startdate><enddate>19870801</enddate><creator>McGillis, Joseph P.</creator><creator>Organist, Michele L.</creator><creator>Payan, Donald G.</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7Z</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19870801</creationdate><title>Immunoaffinity purification of membrane protein constituents of the IM-9 lymphoblast receptor for substance P</title><author>McGillis, Joseph P. ; Organist, Michele L. ; Payan, Donald G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c388t-5f7f5d15a492a0ee03309a51c0aa3d21bd85b035eca9a4b63ea355fa0bd89b803</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1987</creationdate><topic>Affinity Labels</topic><topic>Cell Line</topic><topic>Chromatography, Affinity - methods</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Humans</topic><topic>immunoaffinity purification</topic><topic>lymphocyte</topic><topic>Lymphocytes - analysis</topic><topic>lymphocytes T</topic><topic>man</topic><topic>Membrane Proteins - isolation & purification</topic><topic>Molecular Weight</topic><topic>Receptors, Neurokinin-1</topic><topic>Receptors, Neurotransmitter - isolation & purification</topic><topic>Solubility</topic><topic>substance P</topic><topic>substance P receptor</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>McGillis, Joseph P.</creatorcontrib><creatorcontrib>Organist, Michele L.</creatorcontrib><creatorcontrib>Payan, Donald G.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 1</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>McGillis, Joseph P.</au><au>Organist, Michele L.</au><au>Payan, Donald G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Immunoaffinity purification of membrane protein constituents of the IM-9 lymphoblast receptor for substance P</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>1987-08-01</date><risdate>1987</risdate><volume>164</volume><issue>2</issue><spage>502</spage><epage>513</epage><pages>502-513</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><abstract>Substance P (SP) is an undecapeptide neuromediator that stimulates human T-lymphocyte function by binding to stereospecific membrane receptors. Human IM-9 cultured B-lymphoblasts express approximately 20,000 receptors per cell for [
125I]SP with a
K
d
of 0.3 nM. [
125I]SP was specifically crosslinked by disuccinimidyl suberate to IM-9 cell membrane proteins of 78, 58, and 33 kDa. An indirect immunoaffinity purification procedure has now been developed based on immunoabsorption of detergent-solubilized [
125I]SP-labeled IM-9 cell membrane proteins to anti-SP antibody that was bound to an epoxide ultraffinity high-performance liquid chromatography column, followed by elution in acidic 8
m urea. The 58- and 33-kDa SP-receptor complexes were purified to apparent homogeneity by immunoaffinity chromatography and identified by autoradiography and silver staining of sodium dodecyl sulfate-polyacrylamide gels.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>2823637</pmid><doi>10.1016/0003-2697(87)90525-2</doi><tpages>12</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-2697 |
| ispartof | Analytical biochemistry, 1987-08, Vol.164 (2), p.502-513 |
| issn | 0003-2697 1096-0309 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Affinity Labels Cell Line Chromatography, Affinity - methods Chromatography, High Pressure Liquid - methods Humans immunoaffinity purification lymphocyte Lymphocytes - analysis lymphocytes T man Membrane Proteins - isolation & purification Molecular Weight Receptors, Neurokinin-1 Receptors, Neurotransmitter - isolation & purification Solubility substance P substance P receptor |
| title | Immunoaffinity purification of membrane protein constituents of the IM-9 lymphoblast receptor for substance P |
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