High-level expression in Escherichia coli of the catalytically active flavin domain of corn leaf NADH:nitrate reductase and its comparison to human NADH:cytochrome B5 reductase
Higher plant nitrate reductase can be divided into three functional domains representing its prosthetic groups: 1) flavin; 2) cytochrome b; and 3) Mo-pterin. The flavin domain has been synthesized by heterologous expression in Escherichia coli using a fragment of a corn leaf NADH:nitrate reductase c...
Gespeichert in:
| Veröffentlicht in: | Biochemical and biophysical research communications 1990-05, Vol.168 (3), p.1285-1291 |
|---|---|
| Hauptverfasser: | , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1291 |
|---|---|
| container_issue | 3 |
| container_start_page | 1285 |
| container_title | Biochemical and biophysical research communications |
| container_volume | 168 |
| creator | Hyde, G.E. (Michigan Technological University, Houghton, MI) Campbell, W.H |
| description | Higher plant nitrate reductase can be divided into three functional domains representing its prosthetic groups: 1) flavin; 2) cytochrome b; and 3) Mo-pterin. The flavin domain has been synthesized by heterologous expression in Escherichia coli using a fragment of a corn leaf NADH:nitrate reductase cDNA clone, Zmnr1, which we had previously isolated and sequenced. A Xho2-BamH1 fragment was cut from Zmnr1, containing the sequence for the flavin domain, and ligated in the BamH1 site of expression vector pET3c. When this construct was expressed in E. coli, a 30 kD polypeptide was found to be newly synthesized. The flavin domain was purified to homogeneity using blue Sepharose and shown to have a molecular weight of 30 kD. The recombinant flavin domain has a ferricyanide reductase specific activity of 1000 mumols NADH oxidized/min/mg protein and a visible spectrum virtually identical to that of human NADH:cytochrome b5 reductase. |
| doi_str_mv | 10.1016/0006-291X(90)91168-R |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_79799399</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>79799399</sourcerecordid><originalsourceid>FETCH-LOGICAL-c321t-57e9cf17acd3146c4c20574cb32b8155db8924211027f53efe63506cac4d9f9f3</originalsourceid><addsrcrecordid>eNpFkduKFDEQhoMo6zj6AqKQK9GL1hz6MNm7dV0dYVFYXfAu1KQr25F0Z0zSg_NWPqIZZ1ihoC7q_6pIPkJecPaWM96-Y4y1lVD8x2vF3ijO21V184AsOFOsEpzVD8niPvKYPEnpJ2Oc1606I2eCr1TNVgvyZ-3uhsrjDj3F39uIKbkwUTfRq2QGjM4MDqgJ3tFgaR6QGsjg99kZ8H5PwWS3Q2o97ArThxFKK0kT4kQ9gqVfLj6szyeXI2SkEfvZZEhIYeqpy6kExy1El8rRHOgwjzAdEbPPwQwxjEjfN__Bp-SRBZ_w2akvye3Hq--X6-r666fPlxfXlZGC56rpUBnLOzC9LG82tRGs6WqzkWKz4k3Tb1ZK1IJzJjrbSLTYyoa1BkzdK6usXJJXx73bGH7NmLIeXTLoPUwY5qQ71SklSy1JfQyaGFKKaPU2uhHiXnOmD6L0wYI-WNCK6X-i9E3BXp72z5sR-3voZKbMnx_nFoKGu_JD-vabYrKtpZB_AZIomaI</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>79799399</pqid></control><display><type>article</type><title>High-level expression in Escherichia coli of the catalytically active flavin domain of corn leaf NADH:nitrate reductase and its comparison to human NADH:cytochrome B5 reductase</title><source>MEDLINE</source><source>ScienceDirect Journals (5 years ago - present)</source><creator>Hyde, G.E. (Michigan Technological University, Houghton, MI) ; Campbell, W.H</creator><creatorcontrib>Hyde, G.E. (Michigan Technological University, Houghton, MI) ; Campbell, W.H</creatorcontrib><description>Higher plant nitrate reductase can be divided into three functional domains representing its prosthetic groups: 1) flavin; 2) cytochrome b; and 3) Mo-pterin. The flavin domain has been synthesized by heterologous expression in Escherichia coli using a fragment of a corn leaf NADH:nitrate reductase cDNA clone, Zmnr1, which we had previously isolated and sequenced. A Xho2-BamH1 fragment was cut from Zmnr1, containing the sequence for the flavin domain, and ligated in the BamH1 site of expression vector pET3c. When this construct was expressed in E. coli, a 30 kD polypeptide was found to be newly synthesized. The flavin domain was purified to homogeneity using blue Sepharose and shown to have a molecular weight of 30 kD. The recombinant flavin domain has a ferricyanide reductase specific activity of 1000 mumols NADH oxidized/min/mg protein and a visible spectrum virtually identical to that of human NADH:cytochrome b5 reductase.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/0006-291X(90)91168-R</identifier><identifier>PMID: 2189408</identifier><language>eng</language><publisher>United States</publisher><subject>ACTIVIDAD CATALITICA ; ACTIVIDAD ENZIMATICA ; ACTIVITE CATALYTIQUE ; ACTIVITE ENZYMATIQUE ; Amino Acid Sequence ; Catalysis ; CATALYTIC ACTIVITY ; COENZIMAS ; COENZYME ; COENZYMES ; Cytochrome Reductases - genetics ; Cytochrome-B Reductase ; Electrophoresis, Polyacrylamide Gel ; ENZYMIC ACTIVITY ; Escherichia coli - genetics ; FEUILLE ; Flavins - genetics ; Flavins - isolation & purification ; Gene Expression ; HOJAS ; Humans ; LEAVES ; METALLOPROTEINE ; METALLOPROTEINS ; METALPROTEINAS ; Molecular Sequence Data ; Nitrate Reductase (NADH) ; Nitrate Reductases - genetics ; Nitrate Reductases - metabolism ; NUCLEOTIDE ; NUCLEOTIDES ; NUCLEOTIDOS ; Peptide Biosynthesis ; Peptides - isolation & purification ; PIGMENT ; PIGMENTOS ; PIGMENTS ; Recombinant Proteins ; Sequence Homology, Nucleic Acid ; ZEA MAYS ; Zea mays - enzymology ; Zea mays - genetics</subject><ispartof>Biochemical and biophysical research communications, 1990-05, Vol.168 (3), p.1285-1291</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c321t-57e9cf17acd3146c4c20574cb32b8155db8924211027f53efe63506cac4d9f9f3</citedby><cites>FETCH-LOGICAL-c321t-57e9cf17acd3146c4c20574cb32b8155db8924211027f53efe63506cac4d9f9f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2189408$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hyde, G.E. (Michigan Technological University, Houghton, MI)</creatorcontrib><creatorcontrib>Campbell, W.H</creatorcontrib><title>High-level expression in Escherichia coli of the catalytically active flavin domain of corn leaf NADH:nitrate reductase and its comparison to human NADH:cytochrome B5 reductase</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>Higher plant nitrate reductase can be divided into three functional domains representing its prosthetic groups: 1) flavin; 2) cytochrome b; and 3) Mo-pterin. The flavin domain has been synthesized by heterologous expression in Escherichia coli using a fragment of a corn leaf NADH:nitrate reductase cDNA clone, Zmnr1, which we had previously isolated and sequenced. A Xho2-BamH1 fragment was cut from Zmnr1, containing the sequence for the flavin domain, and ligated in the BamH1 site of expression vector pET3c. When this construct was expressed in E. coli, a 30 kD polypeptide was found to be newly synthesized. The flavin domain was purified to homogeneity using blue Sepharose and shown to have a molecular weight of 30 kD. The recombinant flavin domain has a ferricyanide reductase specific activity of 1000 mumols NADH oxidized/min/mg protein and a visible spectrum virtually identical to that of human NADH:cytochrome b5 reductase.</description><subject>ACTIVIDAD CATALITICA</subject><subject>ACTIVIDAD ENZIMATICA</subject><subject>ACTIVITE CATALYTIQUE</subject><subject>ACTIVITE ENZYMATIQUE</subject><subject>Amino Acid Sequence</subject><subject>Catalysis</subject><subject>CATALYTIC ACTIVITY</subject><subject>COENZIMAS</subject><subject>COENZYME</subject><subject>COENZYMES</subject><subject>Cytochrome Reductases - genetics</subject><subject>Cytochrome-B Reductase</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>ENZYMIC ACTIVITY</subject><subject>Escherichia coli - genetics</subject><subject>FEUILLE</subject><subject>Flavins - genetics</subject><subject>Flavins - isolation & purification</subject><subject>Gene Expression</subject><subject>HOJAS</subject><subject>Humans</subject><subject>LEAVES</subject><subject>METALLOPROTEINE</subject><subject>METALLOPROTEINS</subject><subject>METALPROTEINAS</subject><subject>Molecular Sequence Data</subject><subject>Nitrate Reductase (NADH)</subject><subject>Nitrate Reductases - genetics</subject><subject>Nitrate Reductases - metabolism</subject><subject>NUCLEOTIDE</subject><subject>NUCLEOTIDES</subject><subject>NUCLEOTIDOS</subject><subject>Peptide Biosynthesis</subject><subject>Peptides - isolation & purification</subject><subject>PIGMENT</subject><subject>PIGMENTOS</subject><subject>PIGMENTS</subject><subject>Recombinant Proteins</subject><subject>Sequence Homology, Nucleic Acid</subject><subject>ZEA MAYS</subject><subject>Zea mays - enzymology</subject><subject>Zea mays - genetics</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkduKFDEQhoMo6zj6AqKQK9GL1hz6MNm7dV0dYVFYXfAu1KQr25F0Z0zSg_NWPqIZZ1ihoC7q_6pIPkJecPaWM96-Y4y1lVD8x2vF3ijO21V184AsOFOsEpzVD8niPvKYPEnpJ2Oc1606I2eCr1TNVgvyZ-3uhsrjDj3F39uIKbkwUTfRq2QGjM4MDqgJ3tFgaR6QGsjg99kZ8H5PwWS3Q2o97ArThxFKK0kT4kQ9gqVfLj6szyeXI2SkEfvZZEhIYeqpy6kExy1El8rRHOgwjzAdEbPPwQwxjEjfN__Bp-SRBZ_w2akvye3Hq--X6-r666fPlxfXlZGC56rpUBnLOzC9LG82tRGs6WqzkWKz4k3Tb1ZK1IJzJjrbSLTYyoa1BkzdK6usXJJXx73bGH7NmLIeXTLoPUwY5qQ71SklSy1JfQyaGFKKaPU2uhHiXnOmD6L0wYI-WNCK6X-i9E3BXp72z5sR-3voZKbMnx_nFoKGu_JD-vabYrKtpZB_AZIomaI</recordid><startdate>19900516</startdate><enddate>19900516</enddate><creator>Hyde, G.E. (Michigan Technological University, Houghton, MI)</creator><creator>Campbell, W.H</creator><scope>FBQ</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>7X8</scope></search><sort><creationdate>19900516</creationdate><title>High-level expression in Escherichia coli of the catalytically active flavin domain of corn leaf NADH:nitrate reductase and its comparison to human NADH:cytochrome B5 reductase</title><author>Hyde, G.E. (Michigan Technological University, Houghton, MI) ; Campbell, W.H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c321t-57e9cf17acd3146c4c20574cb32b8155db8924211027f53efe63506cac4d9f9f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>ACTIVIDAD CATALITICA</topic><topic>ACTIVIDAD ENZIMATICA</topic><topic>ACTIVITE CATALYTIQUE</topic><topic>ACTIVITE ENZYMATIQUE</topic><topic>Amino Acid Sequence</topic><topic>Catalysis</topic><topic>CATALYTIC ACTIVITY</topic><topic>COENZIMAS</topic><topic>COENZYME</topic><topic>COENZYMES</topic><topic>Cytochrome Reductases - genetics</topic><topic>Cytochrome-B Reductase</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>ENZYMIC ACTIVITY</topic><topic>Escherichia coli - genetics</topic><topic>FEUILLE</topic><topic>Flavins - genetics</topic><topic>Flavins - isolation & purification</topic><topic>Gene Expression</topic><topic>HOJAS</topic><topic>Humans</topic><topic>LEAVES</topic><topic>METALLOPROTEINE</topic><topic>METALLOPROTEINS</topic><topic>METALPROTEINAS</topic><topic>Molecular Sequence Data</topic><topic>Nitrate Reductase (NADH)</topic><topic>Nitrate Reductases - genetics</topic><topic>Nitrate Reductases - metabolism</topic><topic>NUCLEOTIDE</topic><topic>NUCLEOTIDES</topic><topic>NUCLEOTIDOS</topic><topic>Peptide Biosynthesis</topic><topic>Peptides - isolation & purification</topic><topic>PIGMENT</topic><topic>PIGMENTOS</topic><topic>PIGMENTS</topic><topic>Recombinant Proteins</topic><topic>Sequence Homology, Nucleic Acid</topic><topic>ZEA MAYS</topic><topic>Zea mays - enzymology</topic><topic>Zea mays - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hyde, G.E. (Michigan Technological University, Houghton, MI)</creatorcontrib><creatorcontrib>Campbell, W.H</creatorcontrib><collection>AGRIS</collection><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><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hyde, G.E. (Michigan Technological University, Houghton, MI)</au><au>Campbell, W.H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>High-level expression in Escherichia coli of the catalytically active flavin domain of corn leaf NADH:nitrate reductase and its comparison to human NADH:cytochrome B5 reductase</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>1990-05-16</date><risdate>1990</risdate><volume>168</volume><issue>3</issue><spage>1285</spage><epage>1291</epage><pages>1285-1291</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>Higher plant nitrate reductase can be divided into three functional domains representing its prosthetic groups: 1) flavin; 2) cytochrome b; and 3) Mo-pterin. The flavin domain has been synthesized by heterologous expression in Escherichia coli using a fragment of a corn leaf NADH:nitrate reductase cDNA clone, Zmnr1, which we had previously isolated and sequenced. A Xho2-BamH1 fragment was cut from Zmnr1, containing the sequence for the flavin domain, and ligated in the BamH1 site of expression vector pET3c. When this construct was expressed in E. coli, a 30 kD polypeptide was found to be newly synthesized. The flavin domain was purified to homogeneity using blue Sepharose and shown to have a molecular weight of 30 kD. The recombinant flavin domain has a ferricyanide reductase specific activity of 1000 mumols NADH oxidized/min/mg protein and a visible spectrum virtually identical to that of human NADH:cytochrome b5 reductase.</abstract><cop>United States</cop><pmid>2189408</pmid><doi>10.1016/0006-291X(90)91168-R</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0006-291X |
| ispartof | Biochemical and biophysical research communications, 1990-05, Vol.168 (3), p.1285-1291 |
| issn | 0006-291X 1090-2104 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_79799399 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | ACTIVIDAD CATALITICA ACTIVIDAD ENZIMATICA ACTIVITE CATALYTIQUE ACTIVITE ENZYMATIQUE Amino Acid Sequence Catalysis CATALYTIC ACTIVITY COENZIMAS COENZYME COENZYMES Cytochrome Reductases - genetics Cytochrome-B Reductase Electrophoresis, Polyacrylamide Gel ENZYMIC ACTIVITY Escherichia coli - genetics FEUILLE Flavins - genetics Flavins - isolation & purification Gene Expression HOJAS Humans LEAVES METALLOPROTEINE METALLOPROTEINS METALPROTEINAS Molecular Sequence Data Nitrate Reductase (NADH) Nitrate Reductases - genetics Nitrate Reductases - metabolism NUCLEOTIDE NUCLEOTIDES NUCLEOTIDOS Peptide Biosynthesis Peptides - isolation & purification PIGMENT PIGMENTOS PIGMENTS Recombinant Proteins Sequence Homology, Nucleic Acid ZEA MAYS Zea mays - enzymology Zea mays - genetics |
| title | High-level expression in Escherichia coli of the catalytically active flavin domain of corn leaf NADH:nitrate reductase and its comparison to human NADH:cytochrome B5 reductase |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-08T06%3A15%3A51IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=High-level%20expression%20in%20Escherichia%20coli%20of%20the%20catalytically%20active%20flavin%20domain%20of%20corn%20leaf%20NADH:nitrate%20reductase%20and%20its%20comparison%20to%20human%20NADH:cytochrome%20B5%20reductase&rft.jtitle=Biochemical%20and%20biophysical%20research%20communications&rft.au=Hyde,%20G.E.%20(Michigan%20Technological%20University,%20Houghton,%20MI)&rft.date=1990-05-16&rft.volume=168&rft.issue=3&rft.spage=1285&rft.epage=1291&rft.pages=1285-1291&rft.issn=0006-291X&rft.eissn=1090-2104&rft_id=info:doi/10.1016/0006-291X(90)91168-R&rft_dat=%3Cproquest_cross%3E79799399%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=79799399&rft_id=info:pmid/2189408&rfr_iscdi=true |