The mechanism of adenosine to inosine conversion by the double-stranded RNA unwinding/modifying activity: a high-performance liquid chromatography-mass spectrometry analysis
We have used directly combined high-performance liquid chromatography-mass spectrometry (LC/MS) to examine the mechanism of the reaction catalyzed by the double-stranded RNA unwinding/modifying activity [Bass & Weintraub (1988) Cell 55, 1089-1098]. A double-stranded RNA substrate in which all ad...
Gespeichert in:
| Veröffentlicht in: | Biochemistry (Easton) 1991-12, Vol.30 (49), p.11507-11514 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 11514 |
|---|---|
| container_issue | 49 |
| container_start_page | 11507 |
| container_title | Biochemistry (Easton) |
| container_volume | 30 |
| creator | Polson, Andrew G Crain, Pamela F Pomerantz, Steven C McCloskey, James A Bass, Brenda L |
| description | We have used directly combined high-performance liquid chromatography-mass spectrometry (LC/MS) to examine the mechanism of the reaction catalyzed by the double-stranded RNA unwinding/modifying activity [Bass & Weintraub (1988) Cell 55, 1089-1098]. A double-stranded RNA substrate in which all adenosines were uniformly labeled with 13C was synthesized. An LC/MS analysis of the nucleoside products from the modified, labeled substrate confirmed that adenosine is modified to inosine during the unwinding/modifying reaction. Most importantly, we found that no carbons are exchanged during the reaction. By including H2(18)O in the reaction, we showed that water serves efficiently as the oxygen donor in vitro. These results are consistent with a hydrolytic deamination mechanism and rule out a base replacement mechanism. Although the double-stranded RNA unwinding/modifying activity appears to utilize a catalytic mechanism similar to that of adenosine deaminase, coformycin, a transition-state analogue, will not inhibit the unwinding/modifying activity. |
| doi_str_mv | 10.1021/bi00113a004 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_osti_</sourceid><recordid>TN_cdi_osti_scitechconnect_5488332</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>16147240</sourcerecordid><originalsourceid>FETCH-LOGICAL-a478t-ca8a57a4cb51a6524de8996a8bb4fb9534f67e1a7874db8257623941262368883</originalsourceid><addsrcrecordid>eNqFkctu1TAQhiMEKofCijWSxQIWKNR2fEm6qypuUrmIHthaE8c5cUnsU9sp5KF4R1zlCFggsZrx_J9nRvMXxWOCXxJMyUlrMSakAozZnWJDOMUlaxp-t9hgjEVJG4HvFw9ivMpPhiU7Ko6IZLISzab4uR0MmowewNk4Id8j6Izz0TqDkkf2kGrvbkyI1jvULijlP52f29GUMQVwnenQ5w9naHbfreus251MvrP9kjMEOtkbm5ZTBGiwu6Hcm9D7MIHTBo32erYd0kPwEyS_C7AflnKCGFHcG51y2aSwIHAwLtHGh8W9HsZoHh3icfHl9avt-dvy4uObd-dnFyUwWadSQw1cAtMtJyA4ZZ2pm0ZA3basbxtesV5IQ0DWknVtTbkUtGoYoTmIuq6r4-Lp2tfHZFXUNuUL5Ru4vJPiLBMVzdCzFdoHfz2bmNRkozbjCM74OSpJedVgWv8XJIIwSRnO4IsV1MHHGEyv9sFOEBZFsLq1Wv1ldaafHNrO7WS6P-zqbdbLVbcxmR-_ZQjflJCV5Gr76TIveSkq_v6rup3-fOVBR3Xl55CPHv85-RdyIMIt</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16147240</pqid></control><display><type>article</type><title>The mechanism of adenosine to inosine conversion by the double-stranded RNA unwinding/modifying activity: a high-performance liquid chromatography-mass spectrometry analysis</title><source>MEDLINE</source><source>American Chemical Society Journals</source><creator>Polson, Andrew G ; Crain, Pamela F ; Pomerantz, Steven C ; McCloskey, James A ; Bass, Brenda L</creator><creatorcontrib>Polson, Andrew G ; Crain, Pamela F ; Pomerantz, Steven C ; McCloskey, James A ; Bass, Brenda L</creatorcontrib><description>We have used directly combined high-performance liquid chromatography-mass spectrometry (LC/MS) to examine the mechanism of the reaction catalyzed by the double-stranded RNA unwinding/modifying activity [Bass & Weintraub (1988) Cell 55, 1089-1098]. A double-stranded RNA substrate in which all adenosines were uniformly labeled with 13C was synthesized. An LC/MS analysis of the nucleoside products from the modified, labeled substrate confirmed that adenosine is modified to inosine during the unwinding/modifying reaction. Most importantly, we found that no carbons are exchanged during the reaction. By including H2(18)O in the reaction, we showed that water serves efficiently as the oxygen donor in vitro. These results are consistent with a hydrolytic deamination mechanism and rule out a base replacement mechanism. Although the double-stranded RNA unwinding/modifying activity appears to utilize a catalytic mechanism similar to that of adenosine deaminase, coformycin, a transition-state analogue, will not inhibit the unwinding/modifying activity.</description><identifier>ISSN: 0006-2960</identifier><identifier>EISSN: 1520-4995</identifier><identifier>DOI: 10.1021/bi00113a004</identifier><identifier>PMID: 1747369</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>550201 - Biochemistry- Tracer Techniques ; ADENOSINE ; Adenosine - chemistry ; AROMATICS ; ATP ; AZAARENES ; BASIC BIOLOGICAL SCIENCES ; CARBON 13 ; CARBON ISOTOPES ; CHEMICAL REACTIONS ; CHROMATOGRAPHY ; Coformycin - pharmacology ; DEAMINATION ; ENZYME ACTIVITY ; EVEN-EVEN NUCLEI ; EVEN-ODD NUCLEI ; Gas Chromatography-Mass Spectrometry ; HETEROCYCLIC COMPOUNDS ; Hydrolysis ; INOSINE ; Inosine - chemistry ; ISOTOPES ; LIGHT NUCLEI ; LIQUID COLUMN CHROMATOGRAPHY ; MASS SPECTROSCOPY ; NUCLEI ; Nucleic Acid Conformation ; Nucleic Acid Heteroduplexes - chemistry ; NUCLEIC ACIDS ; NUCLEOSIDES ; NUCLEOTIDES ; ORGANIC COMPOUNDS ; ORGANIC NITROGEN COMPOUNDS ; Oxygen - chemistry ; OXYGEN 18 ; OXYGEN ISOTOPES ; PURINES ; RIBOSIDES ; RNA ; RNA, Double-Stranded - chemistry ; RNA, Double-Stranded - drug effects ; SEPARATION PROCESSES ; SPECTROSCOPY ; STABLE ISOTOPES ; SUBSTRATES ; Water - chemistry</subject><ispartof>Biochemistry (Easton), 1991-12, Vol.30 (49), p.11507-11514</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a478t-ca8a57a4cb51a6524de8996a8bb4fb9534f67e1a7874db8257623941262368883</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/bi00113a004$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/bi00113a004$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>230,315,782,786,887,2767,27083,27931,27932,56745,56795</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1747369$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/5488332$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Polson, Andrew G</creatorcontrib><creatorcontrib>Crain, Pamela F</creatorcontrib><creatorcontrib>Pomerantz, Steven C</creatorcontrib><creatorcontrib>McCloskey, James A</creatorcontrib><creatorcontrib>Bass, Brenda L</creatorcontrib><title>The mechanism of adenosine to inosine conversion by the double-stranded RNA unwinding/modifying activity: a high-performance liquid chromatography-mass spectrometry analysis</title><title>Biochemistry (Easton)</title><addtitle>Biochemistry</addtitle><description>We have used directly combined high-performance liquid chromatography-mass spectrometry (LC/MS) to examine the mechanism of the reaction catalyzed by the double-stranded RNA unwinding/modifying activity [Bass & Weintraub (1988) Cell 55, 1089-1098]. A double-stranded RNA substrate in which all adenosines were uniformly labeled with 13C was synthesized. An LC/MS analysis of the nucleoside products from the modified, labeled substrate confirmed that adenosine is modified to inosine during the unwinding/modifying reaction. Most importantly, we found that no carbons are exchanged during the reaction. By including H2(18)O in the reaction, we showed that water serves efficiently as the oxygen donor in vitro. These results are consistent with a hydrolytic deamination mechanism and rule out a base replacement mechanism. Although the double-stranded RNA unwinding/modifying activity appears to utilize a catalytic mechanism similar to that of adenosine deaminase, coformycin, a transition-state analogue, will not inhibit the unwinding/modifying activity.</description><subject>550201 - Biochemistry- Tracer Techniques</subject><subject>ADENOSINE</subject><subject>Adenosine - chemistry</subject><subject>AROMATICS</subject><subject>ATP</subject><subject>AZAARENES</subject><subject>BASIC BIOLOGICAL SCIENCES</subject><subject>CARBON 13</subject><subject>CARBON ISOTOPES</subject><subject>CHEMICAL REACTIONS</subject><subject>CHROMATOGRAPHY</subject><subject>Coformycin - pharmacology</subject><subject>DEAMINATION</subject><subject>ENZYME ACTIVITY</subject><subject>EVEN-EVEN NUCLEI</subject><subject>EVEN-ODD NUCLEI</subject><subject>Gas Chromatography-Mass Spectrometry</subject><subject>HETEROCYCLIC COMPOUNDS</subject><subject>Hydrolysis</subject><subject>INOSINE</subject><subject>Inosine - chemistry</subject><subject>ISOTOPES</subject><subject>LIGHT NUCLEI</subject><subject>LIQUID COLUMN CHROMATOGRAPHY</subject><subject>MASS SPECTROSCOPY</subject><subject>NUCLEI</subject><subject>Nucleic Acid Conformation</subject><subject>Nucleic Acid Heteroduplexes - chemistry</subject><subject>NUCLEIC ACIDS</subject><subject>NUCLEOSIDES</subject><subject>NUCLEOTIDES</subject><subject>ORGANIC COMPOUNDS</subject><subject>ORGANIC NITROGEN COMPOUNDS</subject><subject>Oxygen - chemistry</subject><subject>OXYGEN 18</subject><subject>OXYGEN ISOTOPES</subject><subject>PURINES</subject><subject>RIBOSIDES</subject><subject>RNA</subject><subject>RNA, Double-Stranded - chemistry</subject><subject>RNA, Double-Stranded - drug effects</subject><subject>SEPARATION PROCESSES</subject><subject>SPECTROSCOPY</subject><subject>STABLE ISOTOPES</subject><subject>SUBSTRATES</subject><subject>Water - chemistry</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkctu1TAQhiMEKofCijWSxQIWKNR2fEm6qypuUrmIHthaE8c5cUnsU9sp5KF4R1zlCFggsZrx_J9nRvMXxWOCXxJMyUlrMSakAozZnWJDOMUlaxp-t9hgjEVJG4HvFw9ivMpPhiU7Ko6IZLISzab4uR0MmowewNk4Id8j6Izz0TqDkkf2kGrvbkyI1jvULijlP52f29GUMQVwnenQ5w9naHbfreus251MvrP9kjMEOtkbm5ZTBGiwu6Hcm9D7MIHTBo32erYd0kPwEyS_C7AflnKCGFHcG51y2aSwIHAwLtHGh8W9HsZoHh3icfHl9avt-dvy4uObd-dnFyUwWadSQw1cAtMtJyA4ZZ2pm0ZA3basbxtesV5IQ0DWknVtTbkUtGoYoTmIuq6r4-Lp2tfHZFXUNuUL5Ru4vJPiLBMVzdCzFdoHfz2bmNRkozbjCM74OSpJedVgWv8XJIIwSRnO4IsV1MHHGEyv9sFOEBZFsLq1Wv1ldaafHNrO7WS6P-zqbdbLVbcxmR-_ZQjflJCV5Gr76TIveSkq_v6rup3-fOVBR3Xl55CPHv85-RdyIMIt</recordid><startdate>19911210</startdate><enddate>19911210</enddate><creator>Polson, Andrew G</creator><creator>Crain, Pamela F</creator><creator>Pomerantz, Steven C</creator><creator>McCloskey, James A</creator><creator>Bass, Brenda L</creator><general>American Chemical Society</general><scope>BSCLL</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>7TM</scope><scope>7X8</scope><scope>OTOTI</scope></search><sort><creationdate>19911210</creationdate><title>The mechanism of adenosine to inosine conversion by the double-stranded RNA unwinding/modifying activity: a high-performance liquid chromatography-mass spectrometry analysis</title><author>Polson, Andrew G ; Crain, Pamela F ; Pomerantz, Steven C ; McCloskey, James A ; Bass, Brenda L</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a478t-ca8a57a4cb51a6524de8996a8bb4fb9534f67e1a7874db8257623941262368883</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>550201 - Biochemistry- Tracer Techniques</topic><topic>ADENOSINE</topic><topic>Adenosine - chemistry</topic><topic>AROMATICS</topic><topic>ATP</topic><topic>AZAARENES</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>CARBON 13</topic><topic>CARBON ISOTOPES</topic><topic>CHEMICAL REACTIONS</topic><topic>CHROMATOGRAPHY</topic><topic>Coformycin - pharmacology</topic><topic>DEAMINATION</topic><topic>ENZYME ACTIVITY</topic><topic>EVEN-EVEN NUCLEI</topic><topic>EVEN-ODD NUCLEI</topic><topic>Gas Chromatography-Mass Spectrometry</topic><topic>HETEROCYCLIC COMPOUNDS</topic><topic>Hydrolysis</topic><topic>INOSINE</topic><topic>Inosine - chemistry</topic><topic>ISOTOPES</topic><topic>LIGHT NUCLEI</topic><topic>LIQUID COLUMN CHROMATOGRAPHY</topic><topic>MASS SPECTROSCOPY</topic><topic>NUCLEI</topic><topic>Nucleic Acid Conformation</topic><topic>Nucleic Acid Heteroduplexes - chemistry</topic><topic>NUCLEIC ACIDS</topic><topic>NUCLEOSIDES</topic><topic>NUCLEOTIDES</topic><topic>ORGANIC COMPOUNDS</topic><topic>ORGANIC NITROGEN COMPOUNDS</topic><topic>Oxygen - chemistry</topic><topic>OXYGEN 18</topic><topic>OXYGEN ISOTOPES</topic><topic>PURINES</topic><topic>RIBOSIDES</topic><topic>RNA</topic><topic>RNA, Double-Stranded - chemistry</topic><topic>RNA, Double-Stranded - drug effects</topic><topic>SEPARATION PROCESSES</topic><topic>SPECTROSCOPY</topic><topic>STABLE ISOTOPES</topic><topic>SUBSTRATES</topic><topic>Water - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Polson, Andrew G</creatorcontrib><creatorcontrib>Crain, Pamela F</creatorcontrib><creatorcontrib>Pomerantz, Steven C</creatorcontrib><creatorcontrib>McCloskey, James A</creatorcontrib><creatorcontrib>Bass, Brenda L</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><jtitle>Biochemistry (Easton)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Polson, Andrew G</au><au>Crain, Pamela F</au><au>Pomerantz, Steven C</au><au>McCloskey, James A</au><au>Bass, Brenda L</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The mechanism of adenosine to inosine conversion by the double-stranded RNA unwinding/modifying activity: a high-performance liquid chromatography-mass spectrometry analysis</atitle><jtitle>Biochemistry (Easton)</jtitle><addtitle>Biochemistry</addtitle><date>1991-12-10</date><risdate>1991</risdate><volume>30</volume><issue>49</issue><spage>11507</spage><epage>11514</epage><pages>11507-11514</pages><issn>0006-2960</issn><eissn>1520-4995</eissn><abstract>We have used directly combined high-performance liquid chromatography-mass spectrometry (LC/MS) to examine the mechanism of the reaction catalyzed by the double-stranded RNA unwinding/modifying activity [Bass & Weintraub (1988) Cell 55, 1089-1098]. A double-stranded RNA substrate in which all adenosines were uniformly labeled with 13C was synthesized. An LC/MS analysis of the nucleoside products from the modified, labeled substrate confirmed that adenosine is modified to inosine during the unwinding/modifying reaction. Most importantly, we found that no carbons are exchanged during the reaction. By including H2(18)O in the reaction, we showed that water serves efficiently as the oxygen donor in vitro. These results are consistent with a hydrolytic deamination mechanism and rule out a base replacement mechanism. Although the double-stranded RNA unwinding/modifying activity appears to utilize a catalytic mechanism similar to that of adenosine deaminase, coformycin, a transition-state analogue, will not inhibit the unwinding/modifying activity.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>1747369</pmid><doi>10.1021/bi00113a004</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0006-2960 |
| ispartof | Biochemistry (Easton), 1991-12, Vol.30 (49), p.11507-11514 |
| issn | 0006-2960 1520-4995 |
| language | eng |
| recordid | cdi_osti_scitechconnect_5488332 |
| source | MEDLINE; American Chemical Society Journals |
| subjects | 550201 - Biochemistry- Tracer Techniques ADENOSINE Adenosine - chemistry AROMATICS ATP AZAARENES BASIC BIOLOGICAL SCIENCES CARBON 13 CARBON ISOTOPES CHEMICAL REACTIONS CHROMATOGRAPHY Coformycin - pharmacology DEAMINATION ENZYME ACTIVITY EVEN-EVEN NUCLEI EVEN-ODD NUCLEI Gas Chromatography-Mass Spectrometry HETEROCYCLIC COMPOUNDS Hydrolysis INOSINE Inosine - chemistry ISOTOPES LIGHT NUCLEI LIQUID COLUMN CHROMATOGRAPHY MASS SPECTROSCOPY NUCLEI Nucleic Acid Conformation Nucleic Acid Heteroduplexes - chemistry NUCLEIC ACIDS NUCLEOSIDES NUCLEOTIDES ORGANIC COMPOUNDS ORGANIC NITROGEN COMPOUNDS Oxygen - chemistry OXYGEN 18 OXYGEN ISOTOPES PURINES RIBOSIDES RNA RNA, Double-Stranded - chemistry RNA, Double-Stranded - drug effects SEPARATION PROCESSES SPECTROSCOPY STABLE ISOTOPES SUBSTRATES Water - chemistry |
| title | The mechanism of adenosine to inosine conversion by the double-stranded RNA unwinding/modifying activity: a high-performance liquid chromatography-mass spectrometry analysis |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-04T19%3A14%3A58IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_osti_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=The%20mechanism%20of%20adenosine%20to%20inosine%20conversion%20by%20the%20double-stranded%20RNA%20unwinding/modifying%20activity:%20a%20high-performance%20liquid%20chromatography-mass%20spectrometry%20analysis&rft.jtitle=Biochemistry%20(Easton)&rft.au=Polson,%20Andrew%20G&rft.date=1991-12-10&rft.volume=30&rft.issue=49&rft.spage=11507&rft.epage=11514&rft.pages=11507-11514&rft.issn=0006-2960&rft.eissn=1520-4995&rft_id=info:doi/10.1021/bi00113a004&rft_dat=%3Cproquest_osti_%3E16147240%3C/proquest_osti_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=16147240&rft_id=info:pmid/1747369&rfr_iscdi=true |