A radiometric kynurenine monooxygenase assay
Kynurenine 3-monooxygenase is a flavin-dependent monooxygenase that catalyzes the oxidation of l-kynurenine to 3-hydroxyl- l-kynurenine in the kynurenine pathway of tryptophan metabolism. The enzyme requires NADH or NADPH as a cofactor. A discontinuous assay that utilizes l-[ 3H]kynurenine as substr...
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| Veröffentlicht in: | Analytical biochemistry 1990, Vol.184 (1), p.55-58 |
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| creator | Wiseman, Jeffrey S. Nichols, James S. |
| description | Kynurenine 3-monooxygenase is a flavin-dependent monooxygenase that catalyzes the oxidation of
l-kynurenine to 3-hydroxyl-
l-kynurenine in the kynurenine pathway of tryptophan metabolism. The enzyme requires NADH or NADPH as a cofactor. A discontinuous assay that utilizes
l-[
3H]kynurenine as substrate is described. The assay offers high precision and a wide range of accessible substrate and cofactor concentrations. The assay was used to measure kinetic isotope effects and the stereospecificity of oxidation of the cofactor. Hydride is transferred from the A-side (pro-
R) of NADH and NADPH since primary deuterium isotope effects were observed for both cofactors when they were deuterated on the A-side but not on the B-side. The large isotope effect on
V
max
K
m
for NADH is sensitive to the concentration of kynurenine, which indicates that NADH can bind before kynurenine. |
| doi_str_mv | 10.1016/0003-2697(90)90010-7 |
| format | Article |
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l-kynurenine to 3-hydroxyl-
l-kynurenine in the kynurenine pathway of tryptophan metabolism. The enzyme requires NADH or NADPH as a cofactor. A discontinuous assay that utilizes
l-[
3H]kynurenine as substrate is described. The assay offers high precision and a wide range of accessible substrate and cofactor concentrations. The assay was used to measure kinetic isotope effects and the stereospecificity of oxidation of the cofactor. Hydride is transferred from the A-side (pro-
R) of NADH and NADPH since primary deuterium isotope effects were observed for both cofactors when they were deuterated on the A-side but not on the B-side. The large isotope effect on
V
max
K
m
for NADH is sensitive to the concentration of kynurenine, which indicates that NADH can bind before kynurenine.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/0003-2697(90)90010-7</identifier><identifier>PMID: 2321759</identifier><identifier>CODEN: ANBCA2</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>AMINO ACIDS ; Analytical, structural and metabolic biochemistry ; ANIMALS ; AROMATICS ; AZAARENES ; AZOLES ; BASIC BIOLOGICAL SCIENCES ; Biological and medical sciences ; BIOLOGICAL PATHWAYS ; CARBOXYLIC ACIDS ; CHROMATOGRAPHY ; COENZYMES ; DEUTERIUM ; ENZYMES ; Enzymes and enzyme inhibitors ; Fundamental and applied biological sciences. Psychology ; HETEROCYCLIC ACIDS ; HETEROCYCLIC COMPOUNDS ; HYDROGEN COMPOUNDS ; HYDROGEN ISOTOPES ; INDOLES ; ISOTOPE APPLICATIONS ; ISOTOPE EFFECTS ; ISOTOPES ; KETO ACIDS ; Kinetics ; KYNURENINE ; Kynurenine - metabolism ; Kynurenine 3-Monooxygenase ; LIGHT NUCLEI ; liver ; Male ; MAMMALS ; MATHEMATICS ; METABOLISM ; Mixed Function Oxygenases - metabolism ; NAD ; NAD - metabolism ; NADP ; NADP - metabolism ; NUCLEI ; NUCLEOTIDES ; ODD-ODD NUCLEI ; ORGANIC ACIDS ; ORGANIC COMPOUNDS ; ORGANIC NITROGEN COMPOUNDS ; OXIDOREDUCTASES ; OXYGENASES ; PYRROLES ; RADIOASSAY ; Radiometry ; RATS ; Rats, Inbred Strains ; REGRESSION ANALYSIS ; RODENTS ; SEPARATION PROCESSES ; STABLE ISOTOPES ; STATISTICS ; TRACER TECHNIQUES ; Tritium ; TRITIUM COMPOUNDS ; TRYPTOPHAN ; TRYPTOPHAN OXYGENASE ; VERTEBRATES 550201 -- Biochemistry-- Tracer Techniques</subject><ispartof>Analytical biochemistry, 1990, Vol.184 (1), p.55-58</ispartof><rights>1990</rights><rights>1990 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c444t-bb4aede86ac0548fa1af3e7533ddd6f4fd3e2f9567d62005a698800b31567fc13</citedby><cites>FETCH-LOGICAL-c444t-bb4aede86ac0548fa1af3e7533ddd6f4fd3e2f9567d62005a698800b31567fc13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0003-2697(90)90010-7$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,780,784,885,3548,4022,27922,27923,27924,45994</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6675060$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2321759$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/6897154$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Wiseman, Jeffrey S.</creatorcontrib><creatorcontrib>Nichols, James S.</creatorcontrib><title>A radiometric kynurenine monooxygenase assay</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>Kynurenine 3-monooxygenase is a flavin-dependent monooxygenase that catalyzes the oxidation of
l-kynurenine to 3-hydroxyl-
l-kynurenine in the kynurenine pathway of tryptophan metabolism. The enzyme requires NADH or NADPH as a cofactor. A discontinuous assay that utilizes
l-[
3H]kynurenine as substrate is described. The assay offers high precision and a wide range of accessible substrate and cofactor concentrations. The assay was used to measure kinetic isotope effects and the stereospecificity of oxidation of the cofactor. Hydride is transferred from the A-side (pro-
R) of NADH and NADPH since primary deuterium isotope effects were observed for both cofactors when they were deuterated on the A-side but not on the B-side. The large isotope effect on
V
max
K
m
for NADH is sensitive to the concentration of kynurenine, which indicates that NADH can bind before kynurenine.</description><subject>AMINO ACIDS</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>ANIMALS</subject><subject>AROMATICS</subject><subject>AZAARENES</subject><subject>AZOLES</subject><subject>BASIC BIOLOGICAL SCIENCES</subject><subject>Biological and medical sciences</subject><subject>BIOLOGICAL PATHWAYS</subject><subject>CARBOXYLIC ACIDS</subject><subject>CHROMATOGRAPHY</subject><subject>COENZYMES</subject><subject>DEUTERIUM</subject><subject>ENZYMES</subject><subject>Enzymes and enzyme inhibitors</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>HETEROCYCLIC ACIDS</subject><subject>HETEROCYCLIC COMPOUNDS</subject><subject>HYDROGEN COMPOUNDS</subject><subject>HYDROGEN ISOTOPES</subject><subject>INDOLES</subject><subject>ISOTOPE APPLICATIONS</subject><subject>ISOTOPE EFFECTS</subject><subject>ISOTOPES</subject><subject>KETO ACIDS</subject><subject>Kinetics</subject><subject>KYNURENINE</subject><subject>Kynurenine - metabolism</subject><subject>Kynurenine 3-Monooxygenase</subject><subject>LIGHT NUCLEI</subject><subject>liver</subject><subject>Male</subject><subject>MAMMALS</subject><subject>MATHEMATICS</subject><subject>METABOLISM</subject><subject>Mixed Function Oxygenases - metabolism</subject><subject>NAD</subject><subject>NAD - metabolism</subject><subject>NADP</subject><subject>NADP - metabolism</subject><subject>NUCLEI</subject><subject>NUCLEOTIDES</subject><subject>ODD-ODD NUCLEI</subject><subject>ORGANIC ACIDS</subject><subject>ORGANIC COMPOUNDS</subject><subject>ORGANIC NITROGEN COMPOUNDS</subject><subject>OXIDOREDUCTASES</subject><subject>OXYGENASES</subject><subject>PYRROLES</subject><subject>RADIOASSAY</subject><subject>Radiometry</subject><subject>RATS</subject><subject>Rats, Inbred Strains</subject><subject>REGRESSION ANALYSIS</subject><subject>RODENTS</subject><subject>SEPARATION PROCESSES</subject><subject>STABLE ISOTOPES</subject><subject>STATISTICS</subject><subject>TRACER TECHNIQUES</subject><subject>Tritium</subject><subject>TRITIUM COMPOUNDS</subject><subject>TRYPTOPHAN</subject><subject>TRYPTOPHAN OXYGENASE</subject><subject>VERTEBRATES 550201 -- Biochemistry-- Tracer Techniques</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkF1rVDEQhkNR6tr2H1RYpIhCj05Ovk5uhFKsCoXe6HXIJhON7klqcla6_96c7rKX9mpg5pmXmYeQcwrvKVD5AQBY10ut3mp4pwEodOqILCho2QED_YwsDsgL8rLWX42hXMhjctyzniqhF-Tyalmsj3nEqUS3_L1Nm4IpJlyOOeX8sP2ByVZc2lrt9pQ8D3Zd8WxfT8j3m0_frr90t3efv15f3XaOcz51qxW36HGQ1oHgQ7DUBoZKMOa9l4EHz7APWkjlZQ8grNTDALBitLWCo-yEvN7l5jpFU12c0P10OSV0k5GDVlTwBr3ZQfcl_9lgncwYq8P12ibMm2qUVqBUT58EW1ivxTA0kO9AV3KtBYO5L3G0ZWsomFm5mX2a2afRYB6VG9XWXu3zN6sR_WFp77jNL_ZzW51dh2KTi_WASakESGjYxx2GzezfiGV-HJNDH8v8t8_x_3f8A-Z5mwg</recordid><startdate>1990</startdate><enddate>1990</enddate><creator>Wiseman, Jeffrey S.</creator><creator>Nichols, James S.</creator><general>Elsevier Inc</general><general>Elsevier</general><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>7QL</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M81</scope><scope>P64</scope><scope>7X8</scope><scope>OTOTI</scope></search><sort><creationdate>1990</creationdate><title>A radiometric kynurenine monooxygenase assay</title><author>Wiseman, Jeffrey S. ; Nichols, James S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c444t-bb4aede86ac0548fa1af3e7533ddd6f4fd3e2f9567d62005a698800b31567fc13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>AMINO ACIDS</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>ANIMALS</topic><topic>AROMATICS</topic><topic>AZAARENES</topic><topic>AZOLES</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>Biological and medical sciences</topic><topic>BIOLOGICAL PATHWAYS</topic><topic>CARBOXYLIC ACIDS</topic><topic>CHROMATOGRAPHY</topic><topic>COENZYMES</topic><topic>DEUTERIUM</topic><topic>ENZYMES</topic><topic>Enzymes and enzyme inhibitors</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>HETEROCYCLIC ACIDS</topic><topic>HETEROCYCLIC COMPOUNDS</topic><topic>HYDROGEN COMPOUNDS</topic><topic>HYDROGEN ISOTOPES</topic><topic>INDOLES</topic><topic>ISOTOPE APPLICATIONS</topic><topic>ISOTOPE EFFECTS</topic><topic>ISOTOPES</topic><topic>KETO ACIDS</topic><topic>Kinetics</topic><topic>KYNURENINE</topic><topic>Kynurenine - metabolism</topic><topic>Kynurenine 3-Monooxygenase</topic><topic>LIGHT NUCLEI</topic><topic>liver</topic><topic>Male</topic><topic>MAMMALS</topic><topic>MATHEMATICS</topic><topic>METABOLISM</topic><topic>Mixed Function Oxygenases - metabolism</topic><topic>NAD</topic><topic>NAD - metabolism</topic><topic>NADP</topic><topic>NADP - metabolism</topic><topic>NUCLEI</topic><topic>NUCLEOTIDES</topic><topic>ODD-ODD NUCLEI</topic><topic>ORGANIC ACIDS</topic><topic>ORGANIC COMPOUNDS</topic><topic>ORGANIC NITROGEN COMPOUNDS</topic><topic>OXIDOREDUCTASES</topic><topic>OXYGENASES</topic><topic>PYRROLES</topic><topic>RADIOASSAY</topic><topic>Radiometry</topic><topic>RATS</topic><topic>Rats, Inbred Strains</topic><topic>REGRESSION ANALYSIS</topic><topic>RODENTS</topic><topic>SEPARATION PROCESSES</topic><topic>STABLE ISOTOPES</topic><topic>STATISTICS</topic><topic>TRACER TECHNIQUES</topic><topic>Tritium</topic><topic>TRITIUM COMPOUNDS</topic><topic>TRYPTOPHAN</topic><topic>TRYPTOPHAN OXYGENASE</topic><topic>VERTEBRATES 550201 -- Biochemistry-- Tracer Techniques</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wiseman, Jeffrey S.</creatorcontrib><creatorcontrib>Nichols, James S.</creatorcontrib><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>Bacteriology Abstracts (Microbiology B)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 3</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wiseman, Jeffrey S.</au><au>Nichols, James S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A radiometric kynurenine monooxygenase assay</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>1990</date><risdate>1990</risdate><volume>184</volume><issue>1</issue><spage>55</spage><epage>58</epage><pages>55-58</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><coden>ANBCA2</coden><abstract>Kynurenine 3-monooxygenase is a flavin-dependent monooxygenase that catalyzes the oxidation of
l-kynurenine to 3-hydroxyl-
l-kynurenine in the kynurenine pathway of tryptophan metabolism. The enzyme requires NADH or NADPH as a cofactor. A discontinuous assay that utilizes
l-[
3H]kynurenine as substrate is described. The assay offers high precision and a wide range of accessible substrate and cofactor concentrations. The assay was used to measure kinetic isotope effects and the stereospecificity of oxidation of the cofactor. Hydride is transferred from the A-side (pro-
R) of NADH and NADPH since primary deuterium isotope effects were observed for both cofactors when they were deuterated on the A-side but not on the B-side. The large isotope effect on
V
max
K
m
for NADH is sensitive to the concentration of kynurenine, which indicates that NADH can bind before kynurenine.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>2321759</pmid><doi>10.1016/0003-2697(90)90010-7</doi><tpages>4</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-2697 |
| ispartof | Analytical biochemistry, 1990, Vol.184 (1), p.55-58 |
| issn | 0003-2697 1096-0309 |
| language | eng |
| recordid | cdi_osti_scitechconnect_6897154 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | AMINO ACIDS Analytical, structural and metabolic biochemistry ANIMALS AROMATICS AZAARENES AZOLES BASIC BIOLOGICAL SCIENCES Biological and medical sciences BIOLOGICAL PATHWAYS CARBOXYLIC ACIDS CHROMATOGRAPHY COENZYMES DEUTERIUM ENZYMES Enzymes and enzyme inhibitors Fundamental and applied biological sciences. Psychology HETEROCYCLIC ACIDS HETEROCYCLIC COMPOUNDS HYDROGEN COMPOUNDS HYDROGEN ISOTOPES INDOLES ISOTOPE APPLICATIONS ISOTOPE EFFECTS ISOTOPES KETO ACIDS Kinetics KYNURENINE Kynurenine - metabolism Kynurenine 3-Monooxygenase LIGHT NUCLEI liver Male MAMMALS MATHEMATICS METABOLISM Mixed Function Oxygenases - metabolism NAD NAD - metabolism NADP NADP - metabolism NUCLEI NUCLEOTIDES ODD-ODD NUCLEI ORGANIC ACIDS ORGANIC COMPOUNDS ORGANIC NITROGEN COMPOUNDS OXIDOREDUCTASES OXYGENASES PYRROLES RADIOASSAY Radiometry RATS Rats, Inbred Strains REGRESSION ANALYSIS RODENTS SEPARATION PROCESSES STABLE ISOTOPES STATISTICS TRACER TECHNIQUES Tritium TRITIUM COMPOUNDS TRYPTOPHAN TRYPTOPHAN OXYGENASE VERTEBRATES 550201 -- Biochemistry-- Tracer Techniques |
| title | A radiometric kynurenine monooxygenase assay |
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