Characterization of a Gi-protein from Trypanosoma cruzi epimastigote membranes
A guanosine 5'-[gamma-[35S]thio]triphosphate-binding activity was detergent-extracted from Trypanosoma cruzi membranes. This binding activity was co-eluted from gel-filtration columns with a factor which, in a heterologous reconstitution system, blocks glucagon stimulation of adenylate cyclase...
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| Veröffentlicht in: | Biochemical journal 1992-10, Vol.287 (2), p.443-446 |
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| container_title | Biochemical journal |
| container_volume | 287 |
| creator | COSO, O. A DIAZ ANEL, A MARTINETTO, H MUSCHIETTI, J. P KAZANIETZ, M FRAIDENRAICH, D TORRES, H. N FLAWIA, M. M |
| description | A guanosine 5'-[gamma-[35S]thio]triphosphate-binding activity was detergent-extracted from Trypanosoma cruzi membranes. This binding activity was co-eluted from gel-filtration columns with a factor which, in a heterologous reconstitution system, blocks glucagon stimulation of adenylate cyclase activity in liver membranes. ADP-ribosylation of these membranes by pertussis toxin eliminated this blocking capacity. Incubation of T. cruzi membranes with activated pertussis toxin and [adenylate-32P]NAD+ led to the incorporation of radioactivity into a labelled product with an apparent M(r) of approx. 43,000. Crude membranes were electrophoresed on SDS/polyacrylamide gels and analysed, by Western blotting, with GA/1 anti-alpha common, AS/7 anti-alpha t, anti-alpha i1 and anti-alpha i2 polyclonal antibodies. These procedures led to the identification of a specific polypeptide band of about 43 kDa. Another polypeptide reacting with the SW/1 anti-beta antibody, of about 30 kDa, was also detected in the membrane fraction. |
| doi_str_mv | 10.1042/bj2870443 |
| format | Article |
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A ; DIAZ ANEL, A ; MARTINETTO, H ; MUSCHIETTI, J. P ; KAZANIETZ, M ; FRAIDENRAICH, D ; TORRES, H. N ; FLAWIA, M. M</creator><creatorcontrib>COSO, O. A ; DIAZ ANEL, A ; MARTINETTO, H ; MUSCHIETTI, J. P ; KAZANIETZ, M ; FRAIDENRAICH, D ; TORRES, H. N ; FLAWIA, M. M</creatorcontrib><description>A guanosine 5'-[gamma-[35S]thio]triphosphate-binding activity was detergent-extracted from Trypanosoma cruzi membranes. This binding activity was co-eluted from gel-filtration columns with a factor which, in a heterologous reconstitution system, blocks glucagon stimulation of adenylate cyclase activity in liver membranes. ADP-ribosylation of these membranes by pertussis toxin eliminated this blocking capacity. Incubation of T. cruzi membranes with activated pertussis toxin and [adenylate-32P]NAD+ led to the incorporation of radioactivity into a labelled product with an apparent M(r) of approx. 43,000. Crude membranes were electrophoresed on SDS/polyacrylamide gels and analysed, by Western blotting, with GA/1 anti-alpha common, AS/7 anti-alpha t, anti-alpha i1 and anti-alpha i2 polyclonal antibodies. These procedures led to the identification of a specific polypeptide band of about 43 kDa. Another polypeptide reacting with the SW/1 anti-beta antibody, of about 30 kDa, was also detected in the membrane fraction.</description><identifier>ISSN: 0264-6021</identifier><identifier>EISSN: 1470-8728</identifier><identifier>DOI: 10.1042/bj2870443</identifier><identifier>PMID: 1445203</identifier><language>eng</language><publisher>Colchester: Portland Press</publisher><subject>Adenosine Diphosphate - metabolism ; Adenylate Cyclase Toxin ; Amino Acid Sequence ; Analytical, structural and metabolic biochemistry ; Animals ; Binding and carrier proteins ; Biological and medical sciences ; Fundamental and applied biological sciences. Psychology ; GTP-Binding Proteins - analysis ; GTP-Binding Proteins - immunology ; GTP-Binding Proteins - physiology ; Guanosine Triphosphate - metabolism ; Macromolecular Substances ; Membrane Proteins - analysis ; Membrane Proteins - immunology ; Membrane Proteins - physiology ; Molecular Sequence Data ; Pertussis Toxin ; Proteins ; Protozoan Proteins - analysis ; Protozoan Proteins - immunology ; Sulfur Radioisotopes ; Trypanosoma cruzi - chemistry ; Trypanosoma cruzi - physiology ; Virulence Factors, Bordetella - pharmacology</subject><ispartof>Biochemical journal, 1992-10, Vol.287 (2), p.443-446</ispartof><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3143-aa22a5f0e5e0ea3d45d6300058f0d20943af2faca82f66c9e5cec77a3ddd9e523</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1133185/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1133185/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4314815$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1445203$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>COSO, O. A</creatorcontrib><creatorcontrib>DIAZ ANEL, A</creatorcontrib><creatorcontrib>MARTINETTO, H</creatorcontrib><creatorcontrib>MUSCHIETTI, J. P</creatorcontrib><creatorcontrib>KAZANIETZ, M</creatorcontrib><creatorcontrib>FRAIDENRAICH, D</creatorcontrib><creatorcontrib>TORRES, H. N</creatorcontrib><creatorcontrib>FLAWIA, M. M</creatorcontrib><title>Characterization of a Gi-protein from Trypanosoma cruzi epimastigote membranes</title><title>Biochemical journal</title><addtitle>Biochem J</addtitle><description>A guanosine 5'-[gamma-[35S]thio]triphosphate-binding activity was detergent-extracted from Trypanosoma cruzi membranes. This binding activity was co-eluted from gel-filtration columns with a factor which, in a heterologous reconstitution system, blocks glucagon stimulation of adenylate cyclase activity in liver membranes. ADP-ribosylation of these membranes by pertussis toxin eliminated this blocking capacity. Incubation of T. cruzi membranes with activated pertussis toxin and [adenylate-32P]NAD+ led to the incorporation of radioactivity into a labelled product with an apparent M(r) of approx. 43,000. Crude membranes were electrophoresed on SDS/polyacrylamide gels and analysed, by Western blotting, with GA/1 anti-alpha common, AS/7 anti-alpha t, anti-alpha i1 and anti-alpha i2 polyclonal antibodies. These procedures led to the identification of a specific polypeptide band of about 43 kDa. Another polypeptide reacting with the SW/1 anti-beta antibody, of about 30 kDa, was also detected in the membrane fraction.</description><subject>Adenosine Diphosphate - metabolism</subject><subject>Adenylate Cyclase Toxin</subject><subject>Amino Acid Sequence</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Binding and carrier proteins</subject><subject>Biological and medical sciences</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>GTP-Binding Proteins - analysis</subject><subject>GTP-Binding Proteins - immunology</subject><subject>GTP-Binding Proteins - physiology</subject><subject>Guanosine Triphosphate - metabolism</subject><subject>Macromolecular Substances</subject><subject>Membrane Proteins - analysis</subject><subject>Membrane Proteins - immunology</subject><subject>Membrane Proteins - physiology</subject><subject>Molecular Sequence Data</subject><subject>Pertussis Toxin</subject><subject>Proteins</subject><subject>Protozoan Proteins - analysis</subject><subject>Protozoan Proteins - immunology</subject><subject>Sulfur Radioisotopes</subject><subject>Trypanosoma cruzi - chemistry</subject><subject>Trypanosoma cruzi - physiology</subject><subject>Virulence Factors, Bordetella - pharmacology</subject><issn>0264-6021</issn><issn>1470-8728</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkMtKAzEUhoMoWi8LH0CYhQguRk9uM9ONIMUbiG50HU4zSY3MTMZkKrRPb0pL1VUI_8d_zvkIOaVwRUGw6-knq0oQgu-QERUl5FXJql0yAlaIvABGD8hhjJ8AVICAfbJPhZAM-Ii8TD4woB5McEscnO8ybzPMHlzeBz8Y12U2-DZ7C4seOx99i5kO86XLTO9ajIObJSprTTsN2Jl4TPYsNtGcbN4j8n5_9zZ5zJ9fH54mt8-55lTwHJExlBaMNGCQ10LWBQcAWVmoGYwFR8ssaqyYLQo9NlIbXZaJrOv0YfyI3Kx7-_m0NbU23RCwUX1IS4WF8ujU_6RzH2rmvxWlnNNKpoKLTUHwX3MTB9W6qE3TpCv8PKqSc1mMq9WkyzWog48xGLsdQkGt5Kut_MSe_d3ql1zbTvn5JseosbFJmXZxi4nkpqKS_wAzk44h</recordid><startdate>19921015</startdate><enddate>19921015</enddate><creator>COSO, O. A</creator><creator>DIAZ ANEL, A</creator><creator>MARTINETTO, H</creator><creator>MUSCHIETTI, J. P</creator><creator>KAZANIETZ, M</creator><creator>FRAIDENRAICH, D</creator><creator>TORRES, H. N</creator><creator>FLAWIA, M. M</creator><general>Portland Press</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>7X8</scope><scope>5PM</scope></search><sort><creationdate>19921015</creationdate><title>Characterization of a Gi-protein from Trypanosoma cruzi epimastigote membranes</title><author>COSO, O. A ; DIAZ ANEL, A ; MARTINETTO, H ; MUSCHIETTI, J. P ; KAZANIETZ, M ; FRAIDENRAICH, D ; TORRES, H. N ; FLAWIA, M. 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Psychology</topic><topic>GTP-Binding Proteins - analysis</topic><topic>GTP-Binding Proteins - immunology</topic><topic>GTP-Binding Proteins - physiology</topic><topic>Guanosine Triphosphate - metabolism</topic><topic>Macromolecular Substances</topic><topic>Membrane Proteins - analysis</topic><topic>Membrane Proteins - immunology</topic><topic>Membrane Proteins - physiology</topic><topic>Molecular Sequence Data</topic><topic>Pertussis Toxin</topic><topic>Proteins</topic><topic>Protozoan Proteins - analysis</topic><topic>Protozoan Proteins - immunology</topic><topic>Sulfur Radioisotopes</topic><topic>Trypanosoma cruzi - chemistry</topic><topic>Trypanosoma cruzi - physiology</topic><topic>Virulence Factors, Bordetella - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>COSO, O. A</creatorcontrib><creatorcontrib>DIAZ ANEL, A</creatorcontrib><creatorcontrib>MARTINETTO, H</creatorcontrib><creatorcontrib>MUSCHIETTI, J. P</creatorcontrib><creatorcontrib>KAZANIETZ, M</creatorcontrib><creatorcontrib>FRAIDENRAICH, D</creatorcontrib><creatorcontrib>TORRES, H. N</creatorcontrib><creatorcontrib>FLAWIA, M. M</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>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Biochemical journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>COSO, O. A</au><au>DIAZ ANEL, A</au><au>MARTINETTO, H</au><au>MUSCHIETTI, J. P</au><au>KAZANIETZ, M</au><au>FRAIDENRAICH, D</au><au>TORRES, H. N</au><au>FLAWIA, M. M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of a Gi-protein from Trypanosoma cruzi epimastigote membranes</atitle><jtitle>Biochemical journal</jtitle><addtitle>Biochem J</addtitle><date>1992-10-15</date><risdate>1992</risdate><volume>287</volume><issue>2</issue><spage>443</spage><epage>446</epage><pages>443-446</pages><issn>0264-6021</issn><eissn>1470-8728</eissn><abstract>A guanosine 5'-[gamma-[35S]thio]triphosphate-binding activity was detergent-extracted from Trypanosoma cruzi membranes. This binding activity was co-eluted from gel-filtration columns with a factor which, in a heterologous reconstitution system, blocks glucagon stimulation of adenylate cyclase activity in liver membranes. ADP-ribosylation of these membranes by pertussis toxin eliminated this blocking capacity. 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| ispartof | Biochemical journal, 1992-10, Vol.287 (2), p.443-446 |
| issn | 0264-6021 1470-8728 |
| language | eng |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Adenosine Diphosphate - metabolism Adenylate Cyclase Toxin Amino Acid Sequence Analytical, structural and metabolic biochemistry Animals Binding and carrier proteins Biological and medical sciences Fundamental and applied biological sciences. Psychology GTP-Binding Proteins - analysis GTP-Binding Proteins - immunology GTP-Binding Proteins - physiology Guanosine Triphosphate - metabolism Macromolecular Substances Membrane Proteins - analysis Membrane Proteins - immunology Membrane Proteins - physiology Molecular Sequence Data Pertussis Toxin Proteins Protozoan Proteins - analysis Protozoan Proteins - immunology Sulfur Radioisotopes Trypanosoma cruzi - chemistry Trypanosoma cruzi - physiology Virulence Factors, Bordetella - pharmacology |
| title | Characterization of a Gi-protein from Trypanosoma cruzi epimastigote membranes |
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