Identification of Cys139 and Glu207 As Catalytically Important Groups in the Active Site of Isopentenyl Diphosphate:Dimethylallyl Diphosphate Isomerase
Isopentenyl diphosphate:dimethylallyl diphosphate isomerase (EC 5.3.3.2) catalyzes the antarafacial [1.3] allylic rearrangement of isopentenyl diphosphate (IPP) to its electrophilic allylic isomer dimethylallyl diphosphate (DMAPP) Active-site thiols at C138 and C139 were recently identified by coval...
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| Veröffentlicht in: | Biochemistry (Easton) 1994-04, Vol.33 (14), p.4212-4217 |
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| creator | Street, Ian P Coffman, Hazel R Poulter, C. Dale Baker, Jonathan A |
| description | Isopentenyl diphosphate:dimethylallyl diphosphate isomerase (EC 5.3.3.2) catalyzes the antarafacial [1.3] allylic rearrangement of isopentenyl diphosphate (IPP) to its electrophilic allylic isomer dimethylallyl diphosphate (DMAPP) Active-site thiols at C138 and C139 were recently identified by covalent modification using active-site-directed irreversible inhibitors [Street, I.P., Poulter, C.D. (1990) Biochemistry 29, 7531-7538; Lu, X.J., Christensen, D. J., and Poulter, C.D. (1992) Biochemistry 31, 9955-9960]. Kinetic studies were conducted with site-directed mutants of IPP isomerase (IPPIase) to evaluate the roles of these amino acids. C138S and C138V mutants were active catalysts with V/K values only-10-fold lower than that of wild-type IPPIase. In contrast,the C139S mutant was a poor catalyst, and the C139A and C139V mutants were inactive. Treatment of the C139S mutant with 3-(fluoromethyl)-3-butenyl diphosphate, an electrophilic active-site-directed irreversible inhibitor, resulted in inactivation of the enzyme by covalent modification of E207. The E207Q and E207V mutants were inactive, suggesting a role for the E207 carboxylate moiety in catalysis |
| doi_str_mv | 10.1021/bi00180a014 |
| format | Article |
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Dale ; Baker, Jonathan A</creator><creatorcontrib>Street, Ian P ; Coffman, Hazel R ; Poulter, C. Dale ; Baker, Jonathan A</creatorcontrib><description>Isopentenyl diphosphate:dimethylallyl diphosphate isomerase (EC 5.3.3.2) catalyzes the antarafacial [1.3] allylic rearrangement of isopentenyl diphosphate (IPP) to its electrophilic allylic isomer dimethylallyl diphosphate (DMAPP) Active-site thiols at C138 and C139 were recently identified by covalent modification using active-site-directed irreversible inhibitors [Street, I.P., Poulter, C.D. (1990) Biochemistry 29, 7531-7538; Lu, X.J., Christensen, D. J., and Poulter, C.D. (1992) Biochemistry 31, 9955-9960]. Kinetic studies were conducted with site-directed mutants of IPP isomerase (IPPIase) to evaluate the roles of these amino acids. C138S and C138V mutants were active catalysts with V/K values only-10-fold lower than that of wild-type IPPIase. In contrast,the C139S mutant was a poor catalyst, and the C139A and C139V mutants were inactive. Treatment of the C139S mutant with 3-(fluoromethyl)-3-butenyl diphosphate, an electrophilic active-site-directed irreversible inhibitor, resulted in inactivation of the enzyme by covalent modification of E207. The E207Q and E207V mutants were inactive, suggesting a role for the E207 carboxylate moiety in catalysis</description><identifier>ISSN: 0006-2960</identifier><identifier>EISSN: 1520-4995</identifier><identifier>DOI: 10.1021/bi00180a014</identifier><identifier>PMID: 7908830</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>ACIDE GLUTAMIQUE ; ACIDO GLUTAMICO ; ACTIVIDAD ENZIMATICA ; ACTIVITE ENZYMATIQUE ; Amino Acid Sequence ; Analytical, structural and metabolic biochemistry ; Base Sequence ; Binding Sites ; Biological and medical sciences ; Carbon-Carbon Double Bond Isomerases ; Catalysis ; CISTEINA ; Claviceps purpurea ; CYSTEINE ; Cysteine - genetics ; Cysteine - metabolism ; DNA Primers ; Enzymes and enzyme inhibitors ; Fundamental and applied biological sciences. Psychology ; Glutamates - genetics ; Glutamates - metabolism ; Glutamic Acid ; ISOMERASAS ; ISOMERASE ; Isomerases ; Isomerases - genetics ; Isomerases - metabolism ; Molecular Sequence Data ; MUTACION ; Mutagenesis, Site-Directed ; MUTATION ; SACCHAROMYCES CEREVISIAE</subject><ispartof>Biochemistry (Easton), 1994-04, Vol.33 (14), p.4212-4217</ispartof><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a499t-c225269df34e6724b23e6f2ea6d5c9a63ad9dfbde19195e84ea351ec7b4bc2653</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/bi00180a014$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/bi00180a014$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4004674$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7908830$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Street, Ian P</creatorcontrib><creatorcontrib>Coffman, Hazel R</creatorcontrib><creatorcontrib>Poulter, C. Dale</creatorcontrib><creatorcontrib>Baker, Jonathan A</creatorcontrib><title>Identification of Cys139 and Glu207 As Catalytically Important Groups in the Active Site of Isopentenyl Diphosphate:Dimethylallyl Diphosphate Isomerase</title><title>Biochemistry (Easton)</title><addtitle>Biochemistry</addtitle><description>Isopentenyl diphosphate:dimethylallyl diphosphate isomerase (EC 5.3.3.2) catalyzes the antarafacial [1.3] allylic rearrangement of isopentenyl diphosphate (IPP) to its electrophilic allylic isomer dimethylallyl diphosphate (DMAPP) Active-site thiols at C138 and C139 were recently identified by covalent modification using active-site-directed irreversible inhibitors [Street, I.P., Poulter, C.D. (1990) Biochemistry 29, 7531-7538; Lu, X.J., Christensen, D. J., and Poulter, C.D. (1992) Biochemistry 31, 9955-9960]. Kinetic studies were conducted with site-directed mutants of IPP isomerase (IPPIase) to evaluate the roles of these amino acids. C138S and C138V mutants were active catalysts with V/K values only-10-fold lower than that of wild-type IPPIase. In contrast,the C139S mutant was a poor catalyst, and the C139A and C139V mutants were inactive. Treatment of the C139S mutant with 3-(fluoromethyl)-3-butenyl diphosphate, an electrophilic active-site-directed irreversible inhibitor, resulted in inactivation of the enzyme by covalent modification of E207. The E207Q and E207V mutants were inactive, suggesting a role for the E207 carboxylate moiety in catalysis</description><subject>ACIDE GLUTAMIQUE</subject><subject>ACIDO GLUTAMICO</subject><subject>ACTIVIDAD ENZIMATICA</subject><subject>ACTIVITE ENZYMATIQUE</subject><subject>Amino Acid Sequence</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Base Sequence</subject><subject>Binding Sites</subject><subject>Biological and medical sciences</subject><subject>Carbon-Carbon Double Bond Isomerases</subject><subject>Catalysis</subject><subject>CISTEINA</subject><subject>Claviceps purpurea</subject><subject>CYSTEINE</subject><subject>Cysteine - genetics</subject><subject>Cysteine - metabolism</subject><subject>DNA Primers</subject><subject>Enzymes and enzyme inhibitors</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glutamates - genetics</subject><subject>Glutamates - metabolism</subject><subject>Glutamic Acid</subject><subject>ISOMERASAS</subject><subject>ISOMERASE</subject><subject>Isomerases</subject><subject>Isomerases - genetics</subject><subject>Isomerases - metabolism</subject><subject>Molecular Sequence Data</subject><subject>MUTACION</subject><subject>Mutagenesis, Site-Directed</subject><subject>MUTATION</subject><subject>SACCHAROMYCES CEREVISIAE</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkUGL1DAYhoMo6-zoyZsg5CB6kGqStunE2zCr4-CA4uyCt_C1_epkbZuapIv9Jf5dM84wKHgK4X3yJHk_Qp5w9pozwd-UhjG-YMB4do_MeC5YkimV3yczxphMhJLsIbn0_jZuM1ZkF-SiUGyxSNmM_NrU2AfTmAqCsT21DV1NnqeKQl_TdTsKVtClpysI0E4hYm070U03WBegD3Tt7Dh4anoa9kiXVTB3SHcm4MG08XaIduynll6ZYW_9sIeAb69Mh2E_tQfXP8nhRIcOPD4iDxpoPT4-rXNy8_7d9epDsv203qyW2wTiF0NSCZELqeomzVAWIitFirIRCLLOKwUyhTqGZY1ccZXjIkNIc45VUWZlJWSezsmLo3dw9seIPujO-ArbFnq0o9dcylSxP-CrI1g5673DRg_OdOAmzZk-jEH_NYZIPztpx7LD-syeeo_581MOPlbaOOgr489YFgcli4MmOWLGB_x5jsF917JIi1xff95puZbZVn39oj9G_umRb8Bq-Oai8mancsbyeOucvDyGUHl9a0fXx2L_-_rfwKm01Q</recordid><startdate>19940412</startdate><enddate>19940412</enddate><creator>Street, Ian P</creator><creator>Coffman, Hazel R</creator><creator>Poulter, C. Dale</creator><creator>Baker, Jonathan A</creator><general>American Chemical Society</general><scope>FBQ</scope><scope>BSCLL</scope><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>M7N</scope></search><sort><creationdate>19940412</creationdate><title>Identification of Cys139 and Glu207 As Catalytically Important Groups in the Active Site of Isopentenyl Diphosphate:Dimethylallyl Diphosphate Isomerase</title><author>Street, Ian P ; Coffman, Hazel R ; Poulter, C. Dale ; Baker, Jonathan A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a499t-c225269df34e6724b23e6f2ea6d5c9a63ad9dfbde19195e84ea351ec7b4bc2653</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>ACIDE GLUTAMIQUE</topic><topic>ACIDO GLUTAMICO</topic><topic>ACTIVIDAD ENZIMATICA</topic><topic>ACTIVITE ENZYMATIQUE</topic><topic>Amino Acid Sequence</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Base Sequence</topic><topic>Binding Sites</topic><topic>Biological and medical sciences</topic><topic>Carbon-Carbon Double Bond Isomerases</topic><topic>Catalysis</topic><topic>CISTEINA</topic><topic>Claviceps purpurea</topic><topic>CYSTEINE</topic><topic>Cysteine - genetics</topic><topic>Cysteine - metabolism</topic><topic>DNA Primers</topic><topic>Enzymes and enzyme inhibitors</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glutamates - genetics</topic><topic>Glutamates - metabolism</topic><topic>Glutamic Acid</topic><topic>ISOMERASAS</topic><topic>ISOMERASE</topic><topic>Isomerases</topic><topic>Isomerases - genetics</topic><topic>Isomerases - metabolism</topic><topic>Molecular Sequence Data</topic><topic>MUTACION</topic><topic>Mutagenesis, Site-Directed</topic><topic>MUTATION</topic><topic>SACCHAROMYCES CEREVISIAE</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Street, Ian P</creatorcontrib><creatorcontrib>Coffman, Hazel R</creatorcontrib><creatorcontrib>Poulter, C. Dale</creatorcontrib><creatorcontrib>Baker, Jonathan A</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><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>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><jtitle>Biochemistry (Easton)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Street, Ian P</au><au>Coffman, Hazel R</au><au>Poulter, C. Dale</au><au>Baker, Jonathan A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of Cys139 and Glu207 As Catalytically Important Groups in the Active Site of Isopentenyl Diphosphate:Dimethylallyl Diphosphate Isomerase</atitle><jtitle>Biochemistry (Easton)</jtitle><addtitle>Biochemistry</addtitle><date>1994-04-12</date><risdate>1994</risdate><volume>33</volume><issue>14</issue><spage>4212</spage><epage>4217</epage><pages>4212-4217</pages><issn>0006-2960</issn><eissn>1520-4995</eissn><abstract>Isopentenyl diphosphate:dimethylallyl diphosphate isomerase (EC 5.3.3.2) catalyzes the antarafacial [1.3] allylic rearrangement of isopentenyl diphosphate (IPP) to its electrophilic allylic isomer dimethylallyl diphosphate (DMAPP) Active-site thiols at C138 and C139 were recently identified by covalent modification using active-site-directed irreversible inhibitors [Street, I.P., Poulter, C.D. (1990) Biochemistry 29, 7531-7538; Lu, X.J., Christensen, D. J., and Poulter, C.D. (1992) Biochemistry 31, 9955-9960]. Kinetic studies were conducted with site-directed mutants of IPP isomerase (IPPIase) to evaluate the roles of these amino acids. C138S and C138V mutants were active catalysts with V/K values only-10-fold lower than that of wild-type IPPIase. In contrast,the C139S mutant was a poor catalyst, and the C139A and C139V mutants were inactive. Treatment of the C139S mutant with 3-(fluoromethyl)-3-butenyl diphosphate, an electrophilic active-site-directed irreversible inhibitor, resulted in inactivation of the enzyme by covalent modification of E207. The E207Q and E207V mutants were inactive, suggesting a role for the E207 carboxylate moiety in catalysis</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>7908830</pmid><doi>10.1021/bi00180a014</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0006-2960 |
| ispartof | Biochemistry (Easton), 1994-04, Vol.33 (14), p.4212-4217 |
| issn | 0006-2960 1520-4995 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_16639065 |
| source | MEDLINE; American Chemical Society Journals |
| subjects | ACIDE GLUTAMIQUE ACIDO GLUTAMICO ACTIVIDAD ENZIMATICA ACTIVITE ENZYMATIQUE Amino Acid Sequence Analytical, structural and metabolic biochemistry Base Sequence Binding Sites Biological and medical sciences Carbon-Carbon Double Bond Isomerases Catalysis CISTEINA Claviceps purpurea CYSTEINE Cysteine - genetics Cysteine - metabolism DNA Primers Enzymes and enzyme inhibitors Fundamental and applied biological sciences. Psychology Glutamates - genetics Glutamates - metabolism Glutamic Acid ISOMERASAS ISOMERASE Isomerases Isomerases - genetics Isomerases - metabolism Molecular Sequence Data MUTACION Mutagenesis, Site-Directed MUTATION SACCHAROMYCES CEREVISIAE |
| title | Identification of Cys139 and Glu207 As Catalytically Important Groups in the Active Site of Isopentenyl Diphosphate:Dimethylallyl Diphosphate Isomerase |
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