Characterization of BciB: A Ferredoxin-Dependent 8‑Vinyl-Protochlorophyllide Reductase from the Green Sulfur Bacterium Chloroherpeton thalassium
Two enzymes, BciA and BciB, are known to reduce the C-8 vinyl group of 8-vinyl protochlorophyllide, producing protochlorophyllide a, during the synthesis of chlorophylls and bacteriochlorophylls in chlorophototrophic bacteria. BciA from the green sulfur bacterium Chlorobaculum tepidum reduces the C-...
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| Veröffentlicht in: | Biochemistry (Easton) 2013-11, Vol.52 (47), p.8442-8451 |
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| creator | Saunders, Allison H Golbeck, John H Bryant, Donald A |
| description | Two enzymes, BciA and BciB, are known to reduce the C-8 vinyl group of 8-vinyl protochlorophyllide, producing protochlorophyllide a, during the synthesis of chlorophylls and bacteriochlorophylls in chlorophototrophic bacteria. BciA from the green sulfur bacterium Chlorobaculum tepidum reduces the C-8 vinyl group using NADPH as the reductant. Cyanobacteria and some other chlorophototrophs have a second, nonhomologous type of 8-vinyl reductase, BciB, but the biochemical properties of this enzyme have not yet been described. In this study, the bciB gene of the green sulfur bacterium Chloroherpeton thalassium was expressed in Escherichia coli, and the recombinant protein was purified and characterized. Recombinant BciB binds a flavin adenine dinucleotide cofactor, and EPR spectroscopy as well as quantitative analyses of bound iron and sulfide suggest that BciB binds two [4Fe–4S] clusters, one of which may not be essential for the activity of the enzyme. Using electrons provided by reduced ferredoxin or dithionite, recombinant BciB was active and reduced the 8-vinyl moiety of the substrate, 8-vinyl protochlorophyllide, producing protochlorophyllide a. A structural model for BciB based on a recent structure for the FrhB subunit of F420-reducing [NiFe]-hydrogenase of Methanothermobacter marburgensis is proposed. Possible reasons for the occurrence and distribution of BciA and BciB among various chlorophototrophs are discussed. |
| doi_str_mv | 10.1021/bi401172b |
| format | Article |
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BciA from the green sulfur bacterium Chlorobaculum tepidum reduces the C-8 vinyl group using NADPH as the reductant. Cyanobacteria and some other chlorophototrophs have a second, nonhomologous type of 8-vinyl reductase, BciB, but the biochemical properties of this enzyme have not yet been described. In this study, the bciB gene of the green sulfur bacterium Chloroherpeton thalassium was expressed in Escherichia coli, and the recombinant protein was purified and characterized. Recombinant BciB binds a flavin adenine dinucleotide cofactor, and EPR spectroscopy as well as quantitative analyses of bound iron and sulfide suggest that BciB binds two [4Fe–4S] clusters, one of which may not be essential for the activity of the enzyme. Using electrons provided by reduced ferredoxin or dithionite, recombinant BciB was active and reduced the 8-vinyl moiety of the substrate, 8-vinyl protochlorophyllide, producing protochlorophyllide a. A structural model for BciB based on a recent structure for the FrhB subunit of F420-reducing [NiFe]-hydrogenase of Methanothermobacter marburgensis is proposed. Possible reasons for the occurrence and distribution of BciA and BciB among various chlorophototrophs are discussed.</description><identifier>ISSN: 0006-2960</identifier><identifier>EISSN: 1520-4995</identifier><identifier>DOI: 10.1021/bi401172b</identifier><identifier>PMID: 24151992</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject><![CDATA[Amino Acid Sequence ; Apoenzymes - chemistry ; Apoenzymes - genetics ; Apoenzymes - isolation & purification ; Apoenzymes - metabolism ; Bacterial Proteins - chemistry ; Bacterial Proteins - genetics ; Bacterial Proteins - isolation & purification ; Bacterial Proteins - metabolism ; Chlorobi - enzymology ; Chlorobi - growth & development ; Cyanobacteria ; Electron Spin Resonance Spectroscopy ; Escherichia coli ; Ferredoxins - metabolism ; Flavin-Adenine Dinucleotide - metabolism ; Iron-Sulfur Proteins - chemistry ; Iron-Sulfur Proteins - genetics ; Iron-Sulfur Proteins - isolation & purification ; Iron-Sulfur Proteins - metabolism ; Isoenzymes - chemistry ; Isoenzymes - genetics ; Isoenzymes - isolation & purification ; Isoenzymes - metabolism ; Metalloporphyrins - metabolism ; Methanothermobacter marburgensis ; Models, Molecular ; Molecular Sequence Data ; Oxidation-Reduction ; Oxidoreductases Acting on CH-CH Group Donors - chemistry ; Oxidoreductases Acting on CH-CH Group Donors - genetics ; Oxidoreductases Acting on CH-CH Group Donors - isolation & purification ; Oxidoreductases Acting on CH-CH Group Donors - metabolism ; Protein Conformation ; Protochlorophyllide - metabolism ; Recombinant Proteins - chemistry ; Recombinant Proteins - isolation & purification ; Recombinant Proteins - metabolism ; Sequence Alignment ; Sequence Homology, Amino Acid ; Substrate Specificity]]></subject><ispartof>Biochemistry (Easton), 2013-11, Vol.52 (47), p.8442-8451</ispartof><rights>Copyright © 2013 American Chemical Society</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a414t-3044d9154e1e758bf7ccdde5f5efeba0d85eec8046870d1a7759feae7d7b8f053</citedby><cites>FETCH-LOGICAL-a414t-3044d9154e1e758bf7ccdde5f5efeba0d85eec8046870d1a7759feae7d7b8f053</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/bi401172b$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/bi401172b$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,776,780,2752,27055,27903,27904,56716,56766</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24151992$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Saunders, Allison H</creatorcontrib><creatorcontrib>Golbeck, John H</creatorcontrib><creatorcontrib>Bryant, Donald A</creatorcontrib><title>Characterization of BciB: A Ferredoxin-Dependent 8‑Vinyl-Protochlorophyllide Reductase from the Green Sulfur Bacterium Chloroherpeton thalassium</title><title>Biochemistry (Easton)</title><addtitle>Biochemistry</addtitle><description>Two enzymes, BciA and BciB, are known to reduce the C-8 vinyl group of 8-vinyl protochlorophyllide, producing protochlorophyllide a, during the synthesis of chlorophylls and bacteriochlorophylls in chlorophototrophic bacteria. BciA from the green sulfur bacterium Chlorobaculum tepidum reduces the C-8 vinyl group using NADPH as the reductant. Cyanobacteria and some other chlorophototrophs have a second, nonhomologous type of 8-vinyl reductase, BciB, but the biochemical properties of this enzyme have not yet been described. In this study, the bciB gene of the green sulfur bacterium Chloroherpeton thalassium was expressed in Escherichia coli, and the recombinant protein was purified and characterized. Recombinant BciB binds a flavin adenine dinucleotide cofactor, and EPR spectroscopy as well as quantitative analyses of bound iron and sulfide suggest that BciB binds two [4Fe–4S] clusters, one of which may not be essential for the activity of the enzyme. Using electrons provided by reduced ferredoxin or dithionite, recombinant BciB was active and reduced the 8-vinyl moiety of the substrate, 8-vinyl protochlorophyllide, producing protochlorophyllide a. A structural model for BciB based on a recent structure for the FrhB subunit of F420-reducing [NiFe]-hydrogenase of Methanothermobacter marburgensis is proposed. Possible reasons for the occurrence and distribution of BciA and BciB among various chlorophototrophs are discussed.</description><subject>Amino Acid Sequence</subject><subject>Apoenzymes - chemistry</subject><subject>Apoenzymes - genetics</subject><subject>Apoenzymes - isolation & purification</subject><subject>Apoenzymes - metabolism</subject><subject>Bacterial Proteins - chemistry</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - isolation & purification</subject><subject>Bacterial Proteins - metabolism</subject><subject>Chlorobi - enzymology</subject><subject>Chlorobi - growth & development</subject><subject>Cyanobacteria</subject><subject>Electron Spin Resonance Spectroscopy</subject><subject>Escherichia coli</subject><subject>Ferredoxins - metabolism</subject><subject>Flavin-Adenine Dinucleotide - metabolism</subject><subject>Iron-Sulfur Proteins - chemistry</subject><subject>Iron-Sulfur Proteins - genetics</subject><subject>Iron-Sulfur Proteins - isolation & purification</subject><subject>Iron-Sulfur Proteins - metabolism</subject><subject>Isoenzymes - chemistry</subject><subject>Isoenzymes - genetics</subject><subject>Isoenzymes - isolation & purification</subject><subject>Isoenzymes - metabolism</subject><subject>Metalloporphyrins - metabolism</subject><subject>Methanothermobacter marburgensis</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Oxidation-Reduction</subject><subject>Oxidoreductases Acting on CH-CH Group Donors - chemistry</subject><subject>Oxidoreductases Acting on CH-CH Group Donors - genetics</subject><subject>Oxidoreductases Acting on CH-CH Group Donors - isolation & purification</subject><subject>Oxidoreductases Acting on CH-CH Group Donors - metabolism</subject><subject>Protein Conformation</subject><subject>Protochlorophyllide - metabolism</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Recombinant Proteins - metabolism</subject><subject>Sequence Alignment</subject><subject>Sequence Homology, Amino Acid</subject><subject>Substrate Specificity</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1O3DAURq2qqEyhi75A5U2ldhFqGztOumOG8iMhgYB2Gzn2tWLkxKntSExXvAJ9xD4JgaGsKnV1Zd3j80n3Q-g9JXuUMPqldZxQKln7Ci2oYKTgdS1eowUhpCxYXZJt9Dalm_nJieRv0DbjVNC6Zgv0e9WpqHSG6H6p7MKAg8VL7ZZf8QE-ghjBhFs3FIcwwmBgyLj6c3f_ww1rX1zEkIPufIhh7NbeOwP4Esyks0qAbQw9zh3g4wgw4KvJ2yni5SZr6vHq6WMHcYQ8x-ZOeZXSvNlFW1b5BO-e5w76fvTtenVSnJ0fn64OzgrFKc_FPuHc1FRwoCBF1VqptTEgrAALrSKmEgC6IrysJDFUSSlqCwqkkW1lidjfQZ823jGGnxOk3PQuafBeDRCm1NDZyvhjzP9RXjJacUbLGf28QXUMKUWwzRhdr-K6oaR5bKt5aWtmPzxrp7YH80L-rWcGPm4ApVNzE6Y4zAf5h-gBjOOfTg</recordid><startdate>20131126</startdate><enddate>20131126</enddate><creator>Saunders, Allison H</creator><creator>Golbeck, John H</creator><creator>Bryant, Donald A</creator><general>American Chemical Society</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>7X8</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20131126</creationdate><title>Characterization of BciB: A Ferredoxin-Dependent 8‑Vinyl-Protochlorophyllide Reductase from the Green Sulfur Bacterium Chloroherpeton thalassium</title><author>Saunders, Allison H ; Golbeck, John H ; Bryant, Donald A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a414t-3044d9154e1e758bf7ccdde5f5efeba0d85eec8046870d1a7759feae7d7b8f053</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Amino Acid Sequence</topic><topic>Apoenzymes - chemistry</topic><topic>Apoenzymes - genetics</topic><topic>Apoenzymes - isolation & purification</topic><topic>Apoenzymes - metabolism</topic><topic>Bacterial Proteins - chemistry</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - isolation & purification</topic><topic>Bacterial Proteins - metabolism</topic><topic>Chlorobi - enzymology</topic><topic>Chlorobi - growth & development</topic><topic>Cyanobacteria</topic><topic>Electron Spin Resonance Spectroscopy</topic><topic>Escherichia coli</topic><topic>Ferredoxins - metabolism</topic><topic>Flavin-Adenine Dinucleotide - metabolism</topic><topic>Iron-Sulfur Proteins - chemistry</topic><topic>Iron-Sulfur Proteins - genetics</topic><topic>Iron-Sulfur Proteins - isolation & purification</topic><topic>Iron-Sulfur Proteins - metabolism</topic><topic>Isoenzymes - chemistry</topic><topic>Isoenzymes - genetics</topic><topic>Isoenzymes - isolation & purification</topic><topic>Isoenzymes - metabolism</topic><topic>Metalloporphyrins - metabolism</topic><topic>Methanothermobacter marburgensis</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Oxidation-Reduction</topic><topic>Oxidoreductases Acting on CH-CH Group Donors - chemistry</topic><topic>Oxidoreductases Acting on CH-CH Group Donors - genetics</topic><topic>Oxidoreductases Acting on CH-CH Group Donors - isolation & purification</topic><topic>Oxidoreductases Acting on CH-CH Group Donors - metabolism</topic><topic>Protein Conformation</topic><topic>Protochlorophyllide - metabolism</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Recombinant Proteins - metabolism</topic><topic>Sequence Alignment</topic><topic>Sequence Homology, Amino Acid</topic><topic>Substrate Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Saunders, Allison H</creatorcontrib><creatorcontrib>Golbeck, John H</creatorcontrib><creatorcontrib>Bryant, Donald A</creatorcontrib><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>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Biochemistry (Easton)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Saunders, Allison H</au><au>Golbeck, John H</au><au>Bryant, Donald A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of BciB: A Ferredoxin-Dependent 8‑Vinyl-Protochlorophyllide Reductase from the Green Sulfur Bacterium Chloroherpeton thalassium</atitle><jtitle>Biochemistry (Easton)</jtitle><addtitle>Biochemistry</addtitle><date>2013-11-26</date><risdate>2013</risdate><volume>52</volume><issue>47</issue><spage>8442</spage><epage>8451</epage><pages>8442-8451</pages><issn>0006-2960</issn><eissn>1520-4995</eissn><abstract>Two enzymes, BciA and BciB, are known to reduce the C-8 vinyl group of 8-vinyl protochlorophyllide, producing protochlorophyllide a, during the synthesis of chlorophylls and bacteriochlorophylls in chlorophototrophic bacteria. BciA from the green sulfur bacterium Chlorobaculum tepidum reduces the C-8 vinyl group using NADPH as the reductant. Cyanobacteria and some other chlorophototrophs have a second, nonhomologous type of 8-vinyl reductase, BciB, but the biochemical properties of this enzyme have not yet been described. In this study, the bciB gene of the green sulfur bacterium Chloroherpeton thalassium was expressed in Escherichia coli, and the recombinant protein was purified and characterized. Recombinant BciB binds a flavin adenine dinucleotide cofactor, and EPR spectroscopy as well as quantitative analyses of bound iron and sulfide suggest that BciB binds two [4Fe–4S] clusters, one of which may not be essential for the activity of the enzyme. Using electrons provided by reduced ferredoxin or dithionite, recombinant BciB was active and reduced the 8-vinyl moiety of the substrate, 8-vinyl protochlorophyllide, producing protochlorophyllide a. A structural model for BciB based on a recent structure for the FrhB subunit of F420-reducing [NiFe]-hydrogenase of Methanothermobacter marburgensis is proposed. Possible reasons for the occurrence and distribution of BciA and BciB among various chlorophototrophs are discussed.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>24151992</pmid><doi>10.1021/bi401172b</doi><tpages>10</tpages></addata></record> |
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| source | ACS Publications; MEDLINE |
| subjects | Amino Acid Sequence Apoenzymes - chemistry Apoenzymes - genetics Apoenzymes - isolation & purification Apoenzymes - metabolism Bacterial Proteins - chemistry Bacterial Proteins - genetics Bacterial Proteins - isolation & purification Bacterial Proteins - metabolism Chlorobi - enzymology Chlorobi - growth & development Cyanobacteria Electron Spin Resonance Spectroscopy Escherichia coli Ferredoxins - metabolism Flavin-Adenine Dinucleotide - metabolism Iron-Sulfur Proteins - chemistry Iron-Sulfur Proteins - genetics Iron-Sulfur Proteins - isolation & purification Iron-Sulfur Proteins - metabolism Isoenzymes - chemistry Isoenzymes - genetics Isoenzymes - isolation & purification Isoenzymes - metabolism Metalloporphyrins - metabolism Methanothermobacter marburgensis Models, Molecular Molecular Sequence Data Oxidation-Reduction Oxidoreductases Acting on CH-CH Group Donors - chemistry Oxidoreductases Acting on CH-CH Group Donors - genetics Oxidoreductases Acting on CH-CH Group Donors - isolation & purification Oxidoreductases Acting on CH-CH Group Donors - metabolism Protein Conformation Protochlorophyllide - metabolism Recombinant Proteins - chemistry Recombinant Proteins - isolation & purification Recombinant Proteins - metabolism Sequence Alignment Sequence Homology, Amino Acid Substrate Specificity |
| title | Characterization of BciB: A Ferredoxin-Dependent 8‑Vinyl-Protochlorophyllide Reductase from the Green Sulfur Bacterium Chloroherpeton thalassium |
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