G protein coupling to the thrombin receptor in Chinese hamster lung fibroblasts
The specific involvement of G proteins in thrombin receptor-mediated Ca2+ mobilization and DNA synthesis has been studied in single Chinese hamster lung fibroblasts (CCL39 cells) activated by the hexapeptide SFLLRN. Immunoblots performed with antibodies directed against the COOH terminus of G protei...
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| Veröffentlicht in: | The Journal of biological chemistry 1994-03, Vol.269 (11), p.8483-8487 |
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| creator | Baffy, G Yang, L Raj, S Manning, D R Williamson, J R |
| description | The specific involvement of G proteins in thrombin receptor-mediated Ca2+ mobilization and DNA synthesis has been studied
in single Chinese hamster lung fibroblasts (CCL39 cells) activated by the hexapeptide SFLLRN. Immunoblots performed with antibodies
directed against the COOH terminus of G protein alpha subunits revealed that alpha q, alpha i, and alpha o were each present
in CCL39 cells. The Ca2+ response to SFLLRN was measured after microinjection of anti-alpha q or anti-alpha o antibodies,
which produced a total blockade in 71 and 46% of cells, respectively. A partial inhibition of the SFLLRN-induced Ca2+ response
was observed in the remaining cells. The lag time between exposure of the cells to SFLLRN and the onset of Ca2+ mobilization
was significantly longer (20-24 s) in cells microinjected with anti-alpha q- or anti-alpha o-antibodies than in control cells
microinjected with preimmune serum (9 +/- 1 s). Moreover, the peak height of the Ca2+ response to SFLLRN was decreased by
36 and 73%, respectively in cells microinjected with anti-alpha q or anti-alpha o antibodies. SFLLRN-induced DNA synthesis
in growth-arrested CCL39 cells was also inhibited (44-78%) by prior microinjection of anti-alpha q or anti-alpha o antibodies.
Anti-alpha 1 antibodies had no effect on the SFLLRN-induced Ca2+ response or on DNA synthesis. These results provide direct
evidence that the thrombin receptor in CCL39 cells is coupled to two different types of G proteins, Gq and Go, both causing
Ca2+ mobilization and mitogenesis. |
| doi_str_mv | 10.1016/s0021-9258(17)37219-8 |
| format | Article |
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in single Chinese hamster lung fibroblasts (CCL39 cells) activated by the hexapeptide SFLLRN. Immunoblots performed with antibodies
directed against the COOH terminus of G protein alpha subunits revealed that alpha q, alpha i, and alpha o were each present
in CCL39 cells. The Ca2+ response to SFLLRN was measured after microinjection of anti-alpha q or anti-alpha o antibodies,
which produced a total blockade in 71 and 46% of cells, respectively. A partial inhibition of the SFLLRN-induced Ca2+ response
was observed in the remaining cells. The lag time between exposure of the cells to SFLLRN and the onset of Ca2+ mobilization
was significantly longer (20-24 s) in cells microinjected with anti-alpha q- or anti-alpha o-antibodies than in control cells
microinjected with preimmune serum (9 +/- 1 s). Moreover, the peak height of the Ca2+ response to SFLLRN was decreased by
36 and 73%, respectively in cells microinjected with anti-alpha q or anti-alpha o antibodies. SFLLRN-induced DNA synthesis
in growth-arrested CCL39 cells was also inhibited (44-78%) by prior microinjection of anti-alpha q or anti-alpha o antibodies.
Anti-alpha 1 antibodies had no effect on the SFLLRN-induced Ca2+ response or on DNA synthesis. These results provide direct
evidence that the thrombin receptor in CCL39 cells is coupled to two different types of G proteins, Gq and Go, both causing
Ca2+ mobilization and mitogenesis.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/s0021-9258(17)37219-8</identifier><identifier>PMID: 8132575</identifier><identifier>CODEN: JBCHA3</identifier><language>eng</language><publisher>Bethesda, MD: American Society for Biochemistry and Molecular Biology</publisher><subject>Amino Acid Sequence ; Animals ; Antibodies ; Biological and medical sciences ; Blotting, Western ; Calcium - metabolism ; Cell Line ; Cell physiology ; Cricetinae ; Cricetulus ; DNA - analysis ; DNA - biosynthesis ; Fibroblasts - drug effects ; Fibroblasts - metabolism ; Fluorescent Dyes ; Fundamental and applied biological sciences. Psychology ; Fura-2 - analogs & derivatives ; GTP-Binding Proteins - analysis ; GTP-Binding Proteins - metabolism ; Lung - metabolism ; Molecular and cellular biology ; Molecular Sequence Data ; Peptide Fragments - immunology ; Peptide Fragments - pharmacology ; Receptors, Cell Surface - drug effects ; Receptors, Cell Surface - metabolism ; Receptors, Thrombin - drug effects ; Receptors, Thrombin - metabolism ; Signal transduction</subject><ispartof>The Journal of biological chemistry, 1994-03, Vol.269 (11), p.8483-8487</ispartof><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c505t-fe6d26c2c06e4da07eab2e43a14b94a25e8031a7dfbd88347df83f6ab8afc3fb3</citedby><cites>FETCH-LOGICAL-c505t-fe6d26c2c06e4da07eab2e43a14b94a25e8031a7dfbd88347df83f6ab8afc3fb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4085205$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8132575$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Baffy, G</creatorcontrib><creatorcontrib>Yang, L</creatorcontrib><creatorcontrib>Raj, S</creatorcontrib><creatorcontrib>Manning, D R</creatorcontrib><creatorcontrib>Williamson, J R</creatorcontrib><title>G protein coupling to the thrombin receptor in Chinese hamster lung fibroblasts</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The specific involvement of G proteins in thrombin receptor-mediated Ca2+ mobilization and DNA synthesis has been studied
in single Chinese hamster lung fibroblasts (CCL39 cells) activated by the hexapeptide SFLLRN. Immunoblots performed with antibodies
directed against the COOH terminus of G protein alpha subunits revealed that alpha q, alpha i, and alpha o were each present
in CCL39 cells. The Ca2+ response to SFLLRN was measured after microinjection of anti-alpha q or anti-alpha o antibodies,
which produced a total blockade in 71 and 46% of cells, respectively. A partial inhibition of the SFLLRN-induced Ca2+ response
was observed in the remaining cells. The lag time between exposure of the cells to SFLLRN and the onset of Ca2+ mobilization
was significantly longer (20-24 s) in cells microinjected with anti-alpha q- or anti-alpha o-antibodies than in control cells
microinjected with preimmune serum (9 +/- 1 s). Moreover, the peak height of the Ca2+ response to SFLLRN was decreased by
36 and 73%, respectively in cells microinjected with anti-alpha q or anti-alpha o antibodies. SFLLRN-induced DNA synthesis
in growth-arrested CCL39 cells was also inhibited (44-78%) by prior microinjection of anti-alpha q or anti-alpha o antibodies.
Anti-alpha 1 antibodies had no effect on the SFLLRN-induced Ca2+ response or on DNA synthesis. These results provide direct
evidence that the thrombin receptor in CCL39 cells is coupled to two different types of G proteins, Gq and Go, both causing
Ca2+ mobilization and mitogenesis.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>Calcium - metabolism</subject><subject>Cell Line</subject><subject>Cell physiology</subject><subject>Cricetinae</subject><subject>Cricetulus</subject><subject>DNA - analysis</subject><subject>DNA - biosynthesis</subject><subject>Fibroblasts - drug effects</subject><subject>Fibroblasts - metabolism</subject><subject>Fluorescent Dyes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fura-2 - analogs & derivatives</subject><subject>GTP-Binding Proteins - analysis</subject><subject>GTP-Binding Proteins - metabolism</subject><subject>Lung - metabolism</subject><subject>Molecular and cellular biology</subject><subject>Molecular Sequence Data</subject><subject>Peptide Fragments - immunology</subject><subject>Peptide Fragments - pharmacology</subject><subject>Receptors, Cell Surface - drug effects</subject><subject>Receptors, Cell Surface - metabolism</subject><subject>Receptors, Thrombin - drug effects</subject><subject>Receptors, Thrombin - metabolism</subject><subject>Signal transduction</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkFGL1DAQx4N4nOvpRzgoKIc-VDNJ06SPspyncHAPKvgWknRyjbTNmrQcfntTd9lXAyED_9_MhB8h10A_AIX2Y6aUQd0xod6BfM8lg65Wz8gOqOI1F_DzOdmdkRfkZc6_aDlNB5fkUgFnQoodebirDikuGObKxfUwhvmxWmK1DFhuipMtQUKHhyWmqtT7IcyYsRrMlBdM1biWBh9sinY0ecmvyIU3Y8bXp_eK_Ph8-33_pb5_uPu6_3RfO0HFUntse9Y65miLTW-oRGMZNtxAY7vGMIGKcjCy97ZXijelUNy3xirjHfeWX5Gb49zy-98r5kVPITscRzNjXLOWLe8kgPovCK1QSjJWQHEEXYo5J_T6kMJk0h8NVG_G9bdNp950apD6n3G9Lbg-LVjthP2566S45G9PucnOjD6Z2YV8xhqqBKMb9uaIDeFxeAoJtQ3RDThp1nYaQKtGcf4XHBaVmQ</recordid><startdate>19940318</startdate><enddate>19940318</enddate><creator>Baffy, G</creator><creator>Yang, L</creator><creator>Raj, S</creator><creator>Manning, D R</creator><creator>Williamson, J R</creator><general>American Society for Biochemistry and Molecular Biology</general><scope>IQODW</scope><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>7QP</scope><scope>7X8</scope></search><sort><creationdate>19940318</creationdate><title>G protein coupling to the thrombin receptor in Chinese hamster lung fibroblasts</title><author>Baffy, G ; Yang, L ; Raj, S ; Manning, D R ; Williamson, J R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c505t-fe6d26c2c06e4da07eab2e43a14b94a25e8031a7dfbd88347df83f6ab8afc3fb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Antibodies</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>Calcium - metabolism</topic><topic>Cell Line</topic><topic>Cell physiology</topic><topic>Cricetinae</topic><topic>Cricetulus</topic><topic>DNA - analysis</topic><topic>DNA - biosynthesis</topic><topic>Fibroblasts - drug effects</topic><topic>Fibroblasts - metabolism</topic><topic>Fluorescent Dyes</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fura-2 - analogs & derivatives</topic><topic>GTP-Binding Proteins - analysis</topic><topic>GTP-Binding Proteins - metabolism</topic><topic>Lung - metabolism</topic><topic>Molecular and cellular biology</topic><topic>Molecular Sequence Data</topic><topic>Peptide Fragments - immunology</topic><topic>Peptide Fragments - pharmacology</topic><topic>Receptors, Cell Surface - drug effects</topic><topic>Receptors, Cell Surface - metabolism</topic><topic>Receptors, Thrombin - drug effects</topic><topic>Receptors, Thrombin - metabolism</topic><topic>Signal transduction</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Baffy, G</creatorcontrib><creatorcontrib>Yang, L</creatorcontrib><creatorcontrib>Raj, S</creatorcontrib><creatorcontrib>Manning, D R</creatorcontrib><creatorcontrib>Williamson, J R</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium & Calcified Tissue 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>Baffy, G</au><au>Yang, L</au><au>Raj, S</au><au>Manning, D R</au><au>Williamson, J R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>G protein coupling to the thrombin receptor in Chinese hamster lung fibroblasts</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1994-03-18</date><risdate>1994</risdate><volume>269</volume><issue>11</issue><spage>8483</spage><epage>8487</epage><pages>8483-8487</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>The specific involvement of G proteins in thrombin receptor-mediated Ca2+ mobilization and DNA synthesis has been studied
in single Chinese hamster lung fibroblasts (CCL39 cells) activated by the hexapeptide SFLLRN. Immunoblots performed with antibodies
directed against the COOH terminus of G protein alpha subunits revealed that alpha q, alpha i, and alpha o were each present
in CCL39 cells. The Ca2+ response to SFLLRN was measured after microinjection of anti-alpha q or anti-alpha o antibodies,
which produced a total blockade in 71 and 46% of cells, respectively. A partial inhibition of the SFLLRN-induced Ca2+ response
was observed in the remaining cells. The lag time between exposure of the cells to SFLLRN and the onset of Ca2+ mobilization
was significantly longer (20-24 s) in cells microinjected with anti-alpha q- or anti-alpha o-antibodies than in control cells
microinjected with preimmune serum (9 +/- 1 s). Moreover, the peak height of the Ca2+ response to SFLLRN was decreased by
36 and 73%, respectively in cells microinjected with anti-alpha q or anti-alpha o antibodies. SFLLRN-induced DNA synthesis
in growth-arrested CCL39 cells was also inhibited (44-78%) by prior microinjection of anti-alpha q or anti-alpha o antibodies.
Anti-alpha 1 antibodies had no effect on the SFLLRN-induced Ca2+ response or on DNA synthesis. These results provide direct
evidence that the thrombin receptor in CCL39 cells is coupled to two different types of G proteins, Gq and Go, both causing
Ca2+ mobilization and mitogenesis.</abstract><cop>Bethesda, MD</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>8132575</pmid><doi>10.1016/s0021-9258(17)37219-8</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
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| ispartof | The Journal of biological chemistry, 1994-03, Vol.269 (11), p.8483-8487 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_76397118 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Amino Acid Sequence Animals Antibodies Biological and medical sciences Blotting, Western Calcium - metabolism Cell Line Cell physiology Cricetinae Cricetulus DNA - analysis DNA - biosynthesis Fibroblasts - drug effects Fibroblasts - metabolism Fluorescent Dyes Fundamental and applied biological sciences. Psychology Fura-2 - analogs & derivatives GTP-Binding Proteins - analysis GTP-Binding Proteins - metabolism Lung - metabolism Molecular and cellular biology Molecular Sequence Data Peptide Fragments - immunology Peptide Fragments - pharmacology Receptors, Cell Surface - drug effects Receptors, Cell Surface - metabolism Receptors, Thrombin - drug effects Receptors, Thrombin - metabolism Signal transduction |
| title | G protein coupling to the thrombin receptor in Chinese hamster lung fibroblasts |
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