Crystal Structure of Bacterial Morphinone Reductase and Properties of the C191A Mutant Enzyme
The crystal structure of the NADH-dependent bacterial flavoenzyme morphinone reductase (MR) has been determined at 2.2-à resolution in complex with the oxidizing substrate codeinone. The structure reveals a dimeric enzyme comprising two 8-fold β/α barrel domains, each bound to FMN, and a subunit...
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| Veröffentlicht in: | The Journal of biological chemistry 2002-08, Vol.277 (34), p.30976-30983 |
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| container_issue | 34 |
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| container_title | The Journal of biological chemistry |
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| creator | Barna, Terez Messiha, Hanan Latif Petosa, Carlo Bruce, Neil C Scrutton, Nigel S Moody, Peter C E |
| description | The crystal structure of the NADH-dependent bacterial flavoenzyme morphinone reductase (MR) has been determined at 2.2-Ã
resolution
in complex with the oxidizing substrate codeinone. The structure reveals a dimeric enzyme comprising two 8-fold β/α barrel
domains, each bound to FMN, and a subunit folding topology and mode of flavin-binding similar to that found in Old Yellow
Enzyme (OYE) and pentaerythritol tetranitrate (PETN) reductase. The subunit interface of MR is formed by interactions from
an N-terminal β strand and helices 2 and 8 of the barrel domain and is different to that seen in OYE. The active site structures
of MR, OYE, and PETN reductase are highly conserved reflecting the ability of these enzymes to catalyze âgenericâ reactions
such as the reduction of 2-cyclohexenone. A region of polypeptide presumed to define the reducing coenzyme specificity is
identified by comparison of the MR structure (NADH-dependent) with that of PETN reductase (NADPH-dependent). The active site
acid identified in OYE (Tyr-196) and conserved in PETN reductase (Tyr-186) is replaced by Cys-191 in MR. Mutagenesis studies
have established that Cys-191 does not act as a crucial acid in the mechanism of reduction of the olefinic bond found in 2-cyclohexenone
and codeinone. |
| doi_str_mv | 10.1074/jbc.M202846200 |
| format | Article |
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resolution
in complex with the oxidizing substrate codeinone. The structure reveals a dimeric enzyme comprising two 8-fold β/α barrel
domains, each bound to FMN, and a subunit folding topology and mode of flavin-binding similar to that found in Old Yellow
Enzyme (OYE) and pentaerythritol tetranitrate (PETN) reductase. The subunit interface of MR is formed by interactions from
an N-terminal β strand and helices 2 and 8 of the barrel domain and is different to that seen in OYE. The active site structures
of MR, OYE, and PETN reductase are highly conserved reflecting the ability of these enzymes to catalyze âgenericâ reactions
such as the reduction of 2-cyclohexenone. A region of polypeptide presumed to define the reducing coenzyme specificity is
identified by comparison of the MR structure (NADH-dependent) with that of PETN reductase (NADPH-dependent). The active site
acid identified in OYE (Tyr-196) and conserved in PETN reductase (Tyr-186) is replaced by Cys-191 in MR. Mutagenesis studies
have established that Cys-191 does not act as a crucial acid in the mechanism of reduction of the olefinic bond found in 2-cyclohexenone
and codeinone.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M202846200</identifier><identifier>PMID: 12048188</identifier><language>eng</language><publisher>United States: American Society for Biochemistry and Molecular Biology</publisher><subject>Bacterial Proteins ; Binding Sites ; Crystallization ; Kinetics ; Mutagenesis ; NADPH Dehydrogenase - chemistry ; Oxidoreductases - chemistry ; Pseudomonas putida - enzymology</subject><ispartof>The Journal of biological chemistry, 2002-08, Vol.277 (34), p.30976-30983</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c457t-20b80264a374ad26caa29eec66c93147d740b1bce3041571822edda34a2743653</citedby><cites>FETCH-LOGICAL-c457t-20b80264a374ad26caa29eec66c93147d740b1bce3041571822edda34a2743653</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12048188$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Barna, Terez</creatorcontrib><creatorcontrib>Messiha, Hanan Latif</creatorcontrib><creatorcontrib>Petosa, Carlo</creatorcontrib><creatorcontrib>Bruce, Neil C</creatorcontrib><creatorcontrib>Scrutton, Nigel S</creatorcontrib><creatorcontrib>Moody, Peter C E</creatorcontrib><title>Crystal Structure of Bacterial Morphinone Reductase and Properties of the C191A Mutant Enzyme</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The crystal structure of the NADH-dependent bacterial flavoenzyme morphinone reductase (MR) has been determined at 2.2-Ã
resolution
in complex with the oxidizing substrate codeinone. The structure reveals a dimeric enzyme comprising two 8-fold β/α barrel
domains, each bound to FMN, and a subunit folding topology and mode of flavin-binding similar to that found in Old Yellow
Enzyme (OYE) and pentaerythritol tetranitrate (PETN) reductase. The subunit interface of MR is formed by interactions from
an N-terminal β strand and helices 2 and 8 of the barrel domain and is different to that seen in OYE. The active site structures
of MR, OYE, and PETN reductase are highly conserved reflecting the ability of these enzymes to catalyze âgenericâ reactions
such as the reduction of 2-cyclohexenone. A region of polypeptide presumed to define the reducing coenzyme specificity is
identified by comparison of the MR structure (NADH-dependent) with that of PETN reductase (NADPH-dependent). The active site
acid identified in OYE (Tyr-196) and conserved in PETN reductase (Tyr-186) is replaced by Cys-191 in MR. Mutagenesis studies
have established that Cys-191 does not act as a crucial acid in the mechanism of reduction of the olefinic bond found in 2-cyclohexenone
and codeinone.</description><subject>Bacterial Proteins</subject><subject>Binding Sites</subject><subject>Crystallization</subject><subject>Kinetics</subject><subject>Mutagenesis</subject><subject>NADPH Dehydrogenase - chemistry</subject><subject>Oxidoreductases - chemistry</subject><subject>Pseudomonas putida - enzymology</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkE1LxDAQhoMouq5ePUoO4q1rJkmb9KiLX7CL4gd4kZCms7aybdckRdZfb2UXnMvA8LzvwEPICbAJMCUvPgs3mXPGtcw4YztkBEyLRKTwtktGjHFIcp7qA3IYwicbRuawTw6AM6lB6xF5n_p1iHZJn6PvXew90m5Br6yL6OvhPO_8qqrbrkX6hOVA2IDUtiV99N0Kfawx_AVihXQKOVzSeR9tG-l1-7Nu8IjsLewy4PF2j8nrzfXL9C6ZPdzeTy9niZOpiglnhWY8k1YoaUueOWt5juiyzOUCpCqVZAUUDgWTkCrQnGNZWiEtV1JkqRiT803vyndfPYZomjo4XC5ti10fDOjhj1QwgJMN6HwXgseFWfm6sX5tgJk_oWYQav6FDoHTbXNfNFj-41uDA3C2Aar6o_quPZqi7lyFjeFKGSGNYLnKxC9_Mnwf</recordid><startdate>20020823</startdate><enddate>20020823</enddate><creator>Barna, Terez</creator><creator>Messiha, Hanan Latif</creator><creator>Petosa, Carlo</creator><creator>Bruce, Neil C</creator><creator>Scrutton, Nigel S</creator><creator>Moody, Peter C E</creator><general>American Society for Biochemistry and Molecular Biology</general><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>7TM</scope><scope>C1K</scope></search><sort><creationdate>20020823</creationdate><title>Crystal Structure of Bacterial Morphinone Reductase and Properties of the C191A Mutant Enzyme</title><author>Barna, Terez ; Messiha, Hanan Latif ; Petosa, Carlo ; Bruce, Neil C ; Scrutton, Nigel S ; Moody, Peter C E</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c457t-20b80264a374ad26caa29eec66c93147d740b1bce3041571822edda34a2743653</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Bacterial Proteins</topic><topic>Binding Sites</topic><topic>Crystallization</topic><topic>Kinetics</topic><topic>Mutagenesis</topic><topic>NADPH Dehydrogenase - chemistry</topic><topic>Oxidoreductases - chemistry</topic><topic>Pseudomonas putida - enzymology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Barna, Terez</creatorcontrib><creatorcontrib>Messiha, Hanan Latif</creatorcontrib><creatorcontrib>Petosa, Carlo</creatorcontrib><creatorcontrib>Bruce, Neil C</creatorcontrib><creatorcontrib>Scrutton, Nigel S</creatorcontrib><creatorcontrib>Moody, Peter C E</creatorcontrib><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>Nucleic Acids Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Barna, Terez</au><au>Messiha, Hanan Latif</au><au>Petosa, Carlo</au><au>Bruce, Neil C</au><au>Scrutton, Nigel S</au><au>Moody, Peter C E</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Crystal Structure of Bacterial Morphinone Reductase and Properties of the C191A Mutant Enzyme</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2002-08-23</date><risdate>2002</risdate><volume>277</volume><issue>34</issue><spage>30976</spage><epage>30983</epage><pages>30976-30983</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>The crystal structure of the NADH-dependent bacterial flavoenzyme morphinone reductase (MR) has been determined at 2.2-Ã
resolution
in complex with the oxidizing substrate codeinone. The structure reveals a dimeric enzyme comprising two 8-fold β/α barrel
domains, each bound to FMN, and a subunit folding topology and mode of flavin-binding similar to that found in Old Yellow
Enzyme (OYE) and pentaerythritol tetranitrate (PETN) reductase. The subunit interface of MR is formed by interactions from
an N-terminal β strand and helices 2 and 8 of the barrel domain and is different to that seen in OYE. The active site structures
of MR, OYE, and PETN reductase are highly conserved reflecting the ability of these enzymes to catalyze âgenericâ reactions
such as the reduction of 2-cyclohexenone. A region of polypeptide presumed to define the reducing coenzyme specificity is
identified by comparison of the MR structure (NADH-dependent) with that of PETN reductase (NADPH-dependent). The active site
acid identified in OYE (Tyr-196) and conserved in PETN reductase (Tyr-186) is replaced by Cys-191 in MR. Mutagenesis studies
have established that Cys-191 does not act as a crucial acid in the mechanism of reduction of the olefinic bond found in 2-cyclohexenone
and codeinone.</abstract><cop>United States</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>12048188</pmid><doi>10.1074/jbc.M202846200</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 2002-08, Vol.277 (34), p.30976-30983 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_18457471 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Bacterial Proteins Binding Sites Crystallization Kinetics Mutagenesis NADPH Dehydrogenase - chemistry Oxidoreductases - chemistry Pseudomonas putida - enzymology |
| title | Crystal Structure of Bacterial Morphinone Reductase and Properties of the C191A Mutant Enzyme |
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