Biochemical evidence for the presence of a single CD3delta and CD3gamma chain in the surface T cell receptor/CD3 complex
The T cell antigen receptor (TCR) consists of an alphabeta heterodimer and associated invariant CD3gamma, delta, epsilon, and zeta chains (TCR/CD3 complex). The general stoichiometry of the receptor complex, which is believed to be one molecule each of TCRalpha, TCRbeta, CD3gamma, and CD3delta and t...
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Veröffentlicht in: | The Journal of biological chemistry 2004-12, Vol.279 (49), p.51068-51074 |
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creator | Pan, Qinghua Gollapudi, Ananta S Dave, Vibhuti P |
description | The T cell antigen receptor (TCR) consists of an alphabeta heterodimer and associated invariant CD3gamma, delta, epsilon, and zeta chains (TCR/CD3 complex). The general stoichiometry of the receptor complex, which is believed to be one molecule each of TCRalpha, TCRbeta, CD3gamma, and CD3delta and two molecules each of CD3epsilon and CD3zeta, is not clearly understood. Although it has been shown that there are two chains of CD3epsilon and CD3zeta, the stoichiometry of CD3gamma or CD3delta chains in the surface antigen receptor complex has not been determined. In the present study, transgenic mice expressing an altered form of mouse CD3delta and CD3gamma were employed to show that the surface TCR complexes contain one molecule each of CD3delta and CD3gamma. Thymocytes from wild type and CD3 chain transgenic mice on the appropriate knockout background were surface-biotinylated and immunoprecipitated using a specific antibody. The immunoprecipitates were resolved in two dimensions under nonreducing/reducing conditions to determine the stoichiometry of CD3delta and CD3gamma in the surface antigen receptor complex. Our data clearly show the presence of one molecule each of CD3delta and CD3gamma in the surface TCR/CD3 complex. |
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The general stoichiometry of the receptor complex, which is believed to be one molecule each of TCRalpha, TCRbeta, CD3gamma, and CD3delta and two molecules each of CD3epsilon and CD3zeta, is not clearly understood. Although it has been shown that there are two chains of CD3epsilon and CD3zeta, the stoichiometry of CD3gamma or CD3delta chains in the surface antigen receptor complex has not been determined. In the present study, transgenic mice expressing an altered form of mouse CD3delta and CD3gamma were employed to show that the surface TCR complexes contain one molecule each of CD3delta and CD3gamma. Thymocytes from wild type and CD3 chain transgenic mice on the appropriate knockout background were surface-biotinylated and immunoprecipitated using a specific antibody. The immunoprecipitates were resolved in two dimensions under nonreducing/reducing conditions to determine the stoichiometry of CD3delta and CD3gamma in the surface antigen receptor complex. Our data clearly show the presence of one molecule each of CD3delta and CD3gamma in the surface TCR/CD3 complex.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>PMID: 15459203</identifier><language>eng</language><publisher>United States</publisher><subject>Animals ; Antibodies, Monoclonal - chemistry ; Biochemistry - methods ; Biotinylation ; Blotting, Western ; CD3 Complex - chemistry ; Cell Membrane - metabolism ; Cell Separation ; Cricetinae ; Flow Cytometry ; Immunoprecipitation ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Mice, Transgenic ; Models, Biological ; Receptors, Antigen, T-Cell - metabolism ; Thymus Gland - metabolism</subject><ispartof>The Journal of biological chemistry, 2004-12, Vol.279 (49), p.51068-51074</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15459203$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pan, Qinghua</creatorcontrib><creatorcontrib>Gollapudi, Ananta S</creatorcontrib><creatorcontrib>Dave, Vibhuti P</creatorcontrib><title>Biochemical evidence for the presence of a single CD3delta and CD3gamma chain in the surface T cell receptor/CD3 complex</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The T cell antigen receptor (TCR) consists of an alphabeta heterodimer and associated invariant CD3gamma, delta, epsilon, and zeta chains (TCR/CD3 complex). The general stoichiometry of the receptor complex, which is believed to be one molecule each of TCRalpha, TCRbeta, CD3gamma, and CD3delta and two molecules each of CD3epsilon and CD3zeta, is not clearly understood. Although it has been shown that there are two chains of CD3epsilon and CD3zeta, the stoichiometry of CD3gamma or CD3delta chains in the surface antigen receptor complex has not been determined. In the present study, transgenic mice expressing an altered form of mouse CD3delta and CD3gamma were employed to show that the surface TCR complexes contain one molecule each of CD3delta and CD3gamma. Thymocytes from wild type and CD3 chain transgenic mice on the appropriate knockout background were surface-biotinylated and immunoprecipitated using a specific antibody. The immunoprecipitates were resolved in two dimensions under nonreducing/reducing conditions to determine the stoichiometry of CD3delta and CD3gamma in the surface antigen receptor complex. Our data clearly show the presence of one molecule each of CD3delta and CD3gamma in the surface TCR/CD3 complex.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - chemistry</subject><subject>Biochemistry - methods</subject><subject>Biotinylation</subject><subject>Blotting, Western</subject><subject>CD3 Complex - chemistry</subject><subject>Cell Membrane - metabolism</subject><subject>Cell Separation</subject><subject>Cricetinae</subject><subject>Flow Cytometry</subject><subject>Immunoprecipitation</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Mice, Knockout</subject><subject>Mice, Transgenic</subject><subject>Models, Biological</subject><subject>Receptors, Antigen, T-Cell - metabolism</subject><subject>Thymus Gland - metabolism</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE9LAzEQxYMotla_guTkbTF_ms3mqNWqUPDSg7clTSZtZLNZk12p396t1rPDg2GG33sMc4KmlFS84IK-naIpIYwWiolqgi5yfidjzRU9RxMq5kIxwqdof--j2UHwRjcYPr2F1gB2MeF-B7hLkH8W0WGNs2-3DeDFA7fQ9Brr1h6GrQ5BY7PTvsWjDr48JKdH2xobaBqcwEDXx3Q70tjE0DWwv0RnTjcZro59htbLx_XiuVi9Pr0s7lZFVwleUODSUEpBaAlQmcowLbjikgDlVklghktHNtYxJZ2RpLRMgQQyp-C043yGbn5juxQ_Bsh9HXw-HKVbiEOuS0mJLFX5L0ilrIQSagSvj-CwCWDrLvmg01f991P-DdEmc08</recordid><startdate>20041203</startdate><enddate>20041203</enddate><creator>Pan, Qinghua</creator><creator>Gollapudi, Ananta S</creator><creator>Dave, Vibhuti P</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7T5</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20041203</creationdate><title>Biochemical evidence for the presence of a single CD3delta and CD3gamma chain in the surface T cell receptor/CD3 complex</title><author>Pan, Qinghua ; Gollapudi, Ananta S ; Dave, Vibhuti P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p853-1e37c111e5a7ee8c8c2a539370e13d97e2c37f0bdf297fc706d29e7e041efaf33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - chemistry</topic><topic>Biochemistry - methods</topic><topic>Biotinylation</topic><topic>Blotting, Western</topic><topic>CD3 Complex - chemistry</topic><topic>Cell Membrane - metabolism</topic><topic>Cell Separation</topic><topic>Cricetinae</topic><topic>Flow Cytometry</topic><topic>Immunoprecipitation</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Mice, Knockout</topic><topic>Mice, Transgenic</topic><topic>Models, Biological</topic><topic>Receptors, Antigen, T-Cell - metabolism</topic><topic>Thymus Gland - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pan, Qinghua</creatorcontrib><creatorcontrib>Gollapudi, Ananta S</creatorcontrib><creatorcontrib>Dave, Vibhuti P</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Immunology Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pan, Qinghua</au><au>Gollapudi, Ananta S</au><au>Dave, Vibhuti P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Biochemical evidence for the presence of a single CD3delta and CD3gamma chain in the surface T cell receptor/CD3 complex</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2004-12-03</date><risdate>2004</risdate><volume>279</volume><issue>49</issue><spage>51068</spage><epage>51074</epage><pages>51068-51074</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>The T cell antigen receptor (TCR) consists of an alphabeta heterodimer and associated invariant CD3gamma, delta, epsilon, and zeta chains (TCR/CD3 complex). The general stoichiometry of the receptor complex, which is believed to be one molecule each of TCRalpha, TCRbeta, CD3gamma, and CD3delta and two molecules each of CD3epsilon and CD3zeta, is not clearly understood. Although it has been shown that there are two chains of CD3epsilon and CD3zeta, the stoichiometry of CD3gamma or CD3delta chains in the surface antigen receptor complex has not been determined. In the present study, transgenic mice expressing an altered form of mouse CD3delta and CD3gamma were employed to show that the surface TCR complexes contain one molecule each of CD3delta and CD3gamma. Thymocytes from wild type and CD3 chain transgenic mice on the appropriate knockout background were surface-biotinylated and immunoprecipitated using a specific antibody. The immunoprecipitates were resolved in two dimensions under nonreducing/reducing conditions to determine the stoichiometry of CD3delta and CD3gamma in the surface antigen receptor complex. Our data clearly show the presence of one molecule each of CD3delta and CD3gamma in the surface TCR/CD3 complex.</abstract><cop>United States</cop><pmid>15459203</pmid><tpages>7</tpages></addata></record> |
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subjects | Animals Antibodies, Monoclonal - chemistry Biochemistry - methods Biotinylation Blotting, Western CD3 Complex - chemistry Cell Membrane - metabolism Cell Separation Cricetinae Flow Cytometry Immunoprecipitation Mice Mice, Inbred C57BL Mice, Knockout Mice, Transgenic Models, Biological Receptors, Antigen, T-Cell - metabolism Thymus Gland - metabolism |
title | Biochemical evidence for the presence of a single CD3delta and CD3gamma chain in the surface T cell receptor/CD3 complex |
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