Membrane-Binding Domain of the Small G Protein G25K Contains an S-(all- Trans-Geranylgeranyl)Cysteine Methyl Ester at its Carboxyl Terminus
We showed previously that a 23-kDa guanine nucleotide-binding protein (G protein) purified from bovine brain membranes is carboxyl methylated and that this modification occurs at or near the membrane-binding domain. In the present study, we identified this small G protein as G25K (formerly termed Gp...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1991-01, Vol.88 (1), p.286-290 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Yamane, Harvey K. Farnsworth, Christopher C. Xie, Hongying Evans, Tony Howald, William N. Gelb, Michael H. Glomset, John A. Clarke, Steven Bernard K.-K. Fung |
| description | We showed previously that a 23-kDa guanine nucleotide-binding protein (G protein) purified from bovine brain membranes is carboxyl methylated and that this modification occurs at or near the membrane-binding domain. In the present study, we identified this small G protein as G25K (formerly termed Gp). We demonstrated that proteolytic digests of3H-methylated G25K contained radiolabeled material that coeluted with synthetic S-(geranylgeranyl)cysteine methyl ester on reversed-phase HPLC. Further treatment by performic acid oxidation yielded radiolabeled material that coeluted with L-cysteic acid methyl ester, verifying that the isoprenoid moiety and carboxyl methyl ester are localized on a C-terminal cysteine residue. Analysis by gas chromatography-coupled mass spectrometry of material released from purified G25K by Raney nickel treatment positively identified the covalently bound lipid as an all-trans-geranylgeranyl (C20) isoprenoid moiety. These results suggest that geranylgeranyl modification and perhaps methyl esterification function in the membrane localization of this small G protein. |
| doi_str_mv | 10.1073/pnas.88.1.286 |
| format | Article |
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Fung</creator><creatorcontrib>Yamane, Harvey K. ; Farnsworth, Christopher C. ; Xie, Hongying ; Evans, Tony ; Howald, William N. ; Gelb, Michael H. ; Glomset, John A. ; Clarke, Steven ; Bernard K.-K. Fung</creatorcontrib><description>We showed previously that a 23-kDa guanine nucleotide-binding protein (G protein) purified from bovine brain membranes is carboxyl methylated and that this modification occurs at or near the membrane-binding domain. In the present study, we identified this small G protein as G25K (formerly termed Gp). We demonstrated that proteolytic digests of3H-methylated G25K contained radiolabeled material that coeluted with synthetic S-(geranylgeranyl)cysteine methyl ester on reversed-phase HPLC. Further treatment by performic acid oxidation yielded radiolabeled material that coeluted with L-cysteic acid methyl ester, verifying that the isoprenoid moiety and carboxyl methyl ester are localized on a C-terminal cysteine residue. Analysis by gas chromatography-coupled mass spectrometry of material released from purified G25K by Raney nickel treatment positively identified the covalently bound lipid as an all-trans-geranylgeranyl (C20) isoprenoid moiety. These results suggest that geranylgeranyl modification and perhaps methyl esterification function in the membrane localization of this small G protein.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.88.1.286</identifier><identifier>PMID: 1898776</identifier><identifier>CODEN: PNASA6</identifier><language>eng</language><publisher>Washington, DC: National Academy of Sciences of the United States of America</publisher><subject>550201 - Biochemistry- Tracer Techniques ; AMINES ; Amino Acid Sequence ; Amino acids ; Analytical, structural and metabolic biochemistry ; ANIMALS ; Antibodies ; AROMATICS ; AZAARENES ; BASIC BIOLOGICAL SCIENCES ; Binding and carrier proteins ; BIOCHEMISTRY ; Biological and medical sciences ; Brain - metabolism ; CATTLE ; Cell Membrane - metabolism ; CHEMICAL REACTIONS ; CHEMISTRY ; CHROMATOGRAPHY ; Cysteine - analogs & derivatives ; Cysteine - analysis ; cysteine methyl esters ; Diterpenes - analysis ; DOMESTIC ANIMALS ; ESTERIFICATION ; ESTERS ; Fundamental and applied biological sciences. Psychology ; GAS CHROMATOGRAPHY ; Gas Chromatography-Mass Spectrometry ; GTP-Binding Proteins - isolation & purification ; GTP-Binding Proteins - metabolism ; GUANINE ; guanine nucleotide-binding protein ; HETEROCYCLIC COMPOUNDS ; HYDROGEN COMPOUNDS ; HYDROXY COMPOUNDS ; MAMMALS ; MASS SPECTROSCOPY ; MEMBRANE PROTEINS ; Methylation ; Molecular Sequence Data ; MOLECULAR STRUCTURE ; Molecular Weight ; Oligopeptides - chemical synthesis ; ORGANIC COMPOUNDS ; ORGANIC NITROGEN COMPOUNDS ; Peptide Fragments - isolation & purification ; Placenta ; Platelets ; PROTEINS ; PURINES ; RUMINANTS ; SEPARATION PROCESSES ; SPECTROSCOPY ; Terpenoids ; TRITIUM COMPOUNDS ; Ungulates ; VERTEBRATES</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1991-01, Vol.88 (1), p.286-290</ispartof><rights>Copyright 1991 The National Academy of Sciences of the United States of America</rights><rights>1991 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c540t-f7c13572ee8cc431b7c6481afbcdebf753f1640c84b46309b759c581fe98baf3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/88/1.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/2355755$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/2355755$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,4009,27901,27902,27903,53768,53770,57994,58227</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19833890$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1898776$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/5043899$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Yamane, Harvey K.</creatorcontrib><creatorcontrib>Farnsworth, Christopher C.</creatorcontrib><creatorcontrib>Xie, Hongying</creatorcontrib><creatorcontrib>Evans, Tony</creatorcontrib><creatorcontrib>Howald, William N.</creatorcontrib><creatorcontrib>Gelb, Michael H.</creatorcontrib><creatorcontrib>Glomset, John A.</creatorcontrib><creatorcontrib>Clarke, Steven</creatorcontrib><creatorcontrib>Bernard K.-K. Fung</creatorcontrib><title>Membrane-Binding Domain of the Small G Protein G25K Contains an S-(all- Trans-Geranylgeranyl)Cysteine Methyl Ester at its Carboxyl Terminus</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>We showed previously that a 23-kDa guanine nucleotide-binding protein (G protein) purified from bovine brain membranes is carboxyl methylated and that this modification occurs at or near the membrane-binding domain. In the present study, we identified this small G protein as G25K (formerly termed Gp). We demonstrated that proteolytic digests of3H-methylated G25K contained radiolabeled material that coeluted with synthetic S-(geranylgeranyl)cysteine methyl ester on reversed-phase HPLC. Further treatment by performic acid oxidation yielded radiolabeled material that coeluted with L-cysteic acid methyl ester, verifying that the isoprenoid moiety and carboxyl methyl ester are localized on a C-terminal cysteine residue. Analysis by gas chromatography-coupled mass spectrometry of material released from purified G25K by Raney nickel treatment positively identified the covalently bound lipid as an all-trans-geranylgeranyl (C20) isoprenoid moiety. These results suggest that geranylgeranyl modification and perhaps methyl esterification function in the membrane localization of this small G protein.</description><subject>550201 - Biochemistry- Tracer Techniques</subject><subject>AMINES</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>ANIMALS</subject><subject>Antibodies</subject><subject>AROMATICS</subject><subject>AZAARENES</subject><subject>BASIC BIOLOGICAL SCIENCES</subject><subject>Binding and carrier proteins</subject><subject>BIOCHEMISTRY</subject><subject>Biological and medical sciences</subject><subject>Brain - metabolism</subject><subject>CATTLE</subject><subject>Cell Membrane - metabolism</subject><subject>CHEMICAL REACTIONS</subject><subject>CHEMISTRY</subject><subject>CHROMATOGRAPHY</subject><subject>Cysteine - analogs & derivatives</subject><subject>Cysteine - analysis</subject><subject>cysteine methyl esters</subject><subject>Diterpenes - analysis</subject><subject>DOMESTIC ANIMALS</subject><subject>ESTERIFICATION</subject><subject>ESTERS</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>GAS CHROMATOGRAPHY</subject><subject>Gas Chromatography-Mass Spectrometry</subject><subject>GTP-Binding Proteins - isolation & purification</subject><subject>GTP-Binding Proteins - metabolism</subject><subject>GUANINE</subject><subject>guanine nucleotide-binding protein</subject><subject>HETEROCYCLIC COMPOUNDS</subject><subject>HYDROGEN COMPOUNDS</subject><subject>HYDROXY COMPOUNDS</subject><subject>MAMMALS</subject><subject>MASS SPECTROSCOPY</subject><subject>MEMBRANE PROTEINS</subject><subject>Methylation</subject><subject>Molecular Sequence Data</subject><subject>MOLECULAR STRUCTURE</subject><subject>Molecular Weight</subject><subject>Oligopeptides - chemical synthesis</subject><subject>ORGANIC COMPOUNDS</subject><subject>ORGANIC NITROGEN COMPOUNDS</subject><subject>Peptide Fragments - isolation & purification</subject><subject>Placenta</subject><subject>Platelets</subject><subject>PROTEINS</subject><subject>PURINES</subject><subject>RUMINANTS</subject><subject>SEPARATION PROCESSES</subject><subject>SPECTROSCOPY</subject><subject>Terpenoids</subject><subject>TRITIUM COMPOUNDS</subject><subject>Ungulates</subject><subject>VERTEBRATES</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkkGPEyEUxydGs9bVozdNiMkaPUyFYRgg8aJ1rcbdaLK9E4ZCy4aBLjBm-xn80tK0bteLnl7g_3v_x-O9qnqO4BRBit9tvExTxqZo2rDuQTVBkKO6azl8WE0gbGjN2qZ9XD1J6RpCyAmDJ9UJYpxR2k2qX5d66KP0uv5o_dL6FfgUBmk9CAbktQZXg3QOzMGPGLIu1_OGfAOz4HNhEpAeXNVvClGDRTFJ9VyXsHWrfXg726ZdlgaXOq-3DpyXYwQyA5sTmMnYh9tyu9BxsH5MT6tHRrqknx3iabX4fL6Yfakvvs-_zj5c1Iq0MNeGKoQJbbRmSrUY9VR1LUPS9Gqpe0MJNqhroWJt33YY8p4SrghDRnPWS4NPq_d7283YD3qptM9ROrGJdpBxK4K04m_F27VYhZ-CQMpJSX-1Tw8pW5GUzVqtVfBeq1yQFjPOC_T6UCOGm1GnLAablHau_HQYk2AQdwQ37L8gIoy3qKMFrPegiiGlqM3dgxEUu0UQu0UQjAkkyiIU_uX9Lo_0fvJFPzvoMinpTBmYsumIcYZLJ7BwLw7czv6PfK_M2T9kYUbnsr7NR5vrlEO8AxtMCCUE_wb9dN3j</recordid><startdate>19910101</startdate><enddate>19910101</enddate><creator>Yamane, Harvey K.</creator><creator>Farnsworth, Christopher C.</creator><creator>Xie, Hongying</creator><creator>Evans, Tony</creator><creator>Howald, William N.</creator><creator>Gelb, Michael H.</creator><creator>Glomset, John A.</creator><creator>Clarke, Steven</creator><creator>Bernard K.-K. Fung</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</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>7QL</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7Z</scope><scope>M81</scope><scope>P64</scope><scope>7X8</scope><scope>OTOTI</scope><scope>5PM</scope></search><sort><creationdate>19910101</creationdate><title>Membrane-Binding Domain of the Small G Protein G25K Contains an S-(all- Trans-Geranylgeranyl)Cysteine Methyl Ester at its Carboxyl Terminus</title><author>Yamane, Harvey K. ; Farnsworth, Christopher C. ; Xie, Hongying ; Evans, Tony ; Howald, William N. ; Gelb, Michael H. ; Glomset, John A. ; Clarke, Steven ; Bernard K.-K. Fung</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c540t-f7c13572ee8cc431b7c6481afbcdebf753f1640c84b46309b759c581fe98baf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>550201 - Biochemistry- Tracer Techniques</topic><topic>AMINES</topic><topic>Amino Acid Sequence</topic><topic>Amino acids</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>ANIMALS</topic><topic>Antibodies</topic><topic>AROMATICS</topic><topic>AZAARENES</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>Binding and carrier proteins</topic><topic>BIOCHEMISTRY</topic><topic>Biological and medical sciences</topic><topic>Brain - metabolism</topic><topic>CATTLE</topic><topic>Cell Membrane - metabolism</topic><topic>CHEMICAL REACTIONS</topic><topic>CHEMISTRY</topic><topic>CHROMATOGRAPHY</topic><topic>Cysteine - analogs & derivatives</topic><topic>Cysteine - analysis</topic><topic>cysteine methyl esters</topic><topic>Diterpenes - analysis</topic><topic>DOMESTIC ANIMALS</topic><topic>ESTERIFICATION</topic><topic>ESTERS</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>GAS CHROMATOGRAPHY</topic><topic>Gas Chromatography-Mass Spectrometry</topic><topic>GTP-Binding Proteins - isolation & purification</topic><topic>GTP-Binding Proteins - metabolism</topic><topic>GUANINE</topic><topic>guanine nucleotide-binding protein</topic><topic>HETEROCYCLIC COMPOUNDS</topic><topic>HYDROGEN COMPOUNDS</topic><topic>HYDROXY COMPOUNDS</topic><topic>MAMMALS</topic><topic>MASS SPECTROSCOPY</topic><topic>MEMBRANE PROTEINS</topic><topic>Methylation</topic><topic>Molecular Sequence Data</topic><topic>MOLECULAR STRUCTURE</topic><topic>Molecular Weight</topic><topic>Oligopeptides - chemical synthesis</topic><topic>ORGANIC COMPOUNDS</topic><topic>ORGANIC NITROGEN COMPOUNDS</topic><topic>Peptide Fragments - isolation & purification</topic><topic>Placenta</topic><topic>Platelets</topic><topic>PROTEINS</topic><topic>PURINES</topic><topic>RUMINANTS</topic><topic>SEPARATION PROCESSES</topic><topic>SPECTROSCOPY</topic><topic>Terpenoids</topic><topic>TRITIUM COMPOUNDS</topic><topic>Ungulates</topic><topic>VERTEBRATES</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yamane, Harvey K.</creatorcontrib><creatorcontrib>Farnsworth, Christopher C.</creatorcontrib><creatorcontrib>Xie, Hongying</creatorcontrib><creatorcontrib>Evans, Tony</creatorcontrib><creatorcontrib>Howald, William N.</creatorcontrib><creatorcontrib>Gelb, Michael H.</creatorcontrib><creatorcontrib>Glomset, John A.</creatorcontrib><creatorcontrib>Clarke, Steven</creatorcontrib><creatorcontrib>Bernard K.-K. Fung</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>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>Biochemistry Abstracts 3</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yamane, Harvey K.</au><au>Farnsworth, Christopher C.</au><au>Xie, Hongying</au><au>Evans, Tony</au><au>Howald, William N.</au><au>Gelb, Michael H.</au><au>Glomset, John A.</au><au>Clarke, Steven</au><au>Bernard K.-K. Fung</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Membrane-Binding Domain of the Small G Protein G25K Contains an S-(all- Trans-Geranylgeranyl)Cysteine Methyl Ester at its Carboxyl Terminus</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1991-01-01</date><risdate>1991</risdate><volume>88</volume><issue>1</issue><spage>286</spage><epage>290</epage><pages>286-290</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><coden>PNASA6</coden><abstract>We showed previously that a 23-kDa guanine nucleotide-binding protein (G protein) purified from bovine brain membranes is carboxyl methylated and that this modification occurs at or near the membrane-binding domain. In the present study, we identified this small G protein as G25K (formerly termed Gp). We demonstrated that proteolytic digests of3H-methylated G25K contained radiolabeled material that coeluted with synthetic S-(geranylgeranyl)cysteine methyl ester on reversed-phase HPLC. Further treatment by performic acid oxidation yielded radiolabeled material that coeluted with L-cysteic acid methyl ester, verifying that the isoprenoid moiety and carboxyl methyl ester are localized on a C-terminal cysteine residue. Analysis by gas chromatography-coupled mass spectrometry of material released from purified G25K by Raney nickel treatment positively identified the covalently bound lipid as an all-trans-geranylgeranyl (C20) isoprenoid moiety. These results suggest that geranylgeranyl modification and perhaps methyl esterification function in the membrane localization of this small G protein.</abstract><cop>Washington, DC</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>1898776</pmid><doi>10.1073/pnas.88.1.286</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1991-01, Vol.88 (1), p.286-290 |
| issn | 0027-8424 1091-6490 |
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| recordid | cdi_proquest_miscellaneous_80365328 |
| source | Jstor Complete Legacy; MEDLINE; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | 550201 - Biochemistry- Tracer Techniques AMINES Amino Acid Sequence Amino acids Analytical, structural and metabolic biochemistry ANIMALS Antibodies AROMATICS AZAARENES BASIC BIOLOGICAL SCIENCES Binding and carrier proteins BIOCHEMISTRY Biological and medical sciences Brain - metabolism CATTLE Cell Membrane - metabolism CHEMICAL REACTIONS CHEMISTRY CHROMATOGRAPHY Cysteine - analogs & derivatives Cysteine - analysis cysteine methyl esters Diterpenes - analysis DOMESTIC ANIMALS ESTERIFICATION ESTERS Fundamental and applied biological sciences. Psychology GAS CHROMATOGRAPHY Gas Chromatography-Mass Spectrometry GTP-Binding Proteins - isolation & purification GTP-Binding Proteins - metabolism GUANINE guanine nucleotide-binding protein HETEROCYCLIC COMPOUNDS HYDROGEN COMPOUNDS HYDROXY COMPOUNDS MAMMALS MASS SPECTROSCOPY MEMBRANE PROTEINS Methylation Molecular Sequence Data MOLECULAR STRUCTURE Molecular Weight Oligopeptides - chemical synthesis ORGANIC COMPOUNDS ORGANIC NITROGEN COMPOUNDS Peptide Fragments - isolation & purification Placenta Platelets PROTEINS PURINES RUMINANTS SEPARATION PROCESSES SPECTROSCOPY Terpenoids TRITIUM COMPOUNDS Ungulates VERTEBRATES |
| title | Membrane-Binding Domain of the Small G Protein G25K Contains an S-(all- Trans-Geranylgeranyl)Cysteine Methyl Ester at its Carboxyl Terminus |
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