Determination of microsomal lauric acid hydroxylase activity by HPLC with flow-through radiochemical quantitation
Identifieur interne : 001540 ( Main/Exploration ); précédent : 001539; suivant : 001541Determination of microsomal lauric acid hydroxylase activity by HPLC with flow-through radiochemical quantitation
Auteurs : Maria C. Romano [États-Unis] ; Kenneth M. Straub [États-Unis] ; Lee Ann P. Yodis [États-Unis] ; Regina D. Eckardt [États-Unis] ; John F. Newton [États-Unis]Source :
- Analytical Biochemistry [ 0003-2697 ] ; 1988.
English descriptors
- KwdEn :
- Teeft :
- Academic press, Assay, Biol, Chem, Control values, Cytochrome, Different isozymes, Enzyme activities, Ether, Flow cell size, Flow rate, Glucose, Hepatic, Hepatic microsomes, Hplc, Hplc effluent ratio, Hydrogen peroxide, Hydroxylase, Hydroxylase activities, Hydroxylases, Hydroxylated metabolites, Hydroxylation, Incubation, Isozymes, Kupfer, Lauric, Lauric acid, Lauric acid metabolites, Metabolism, Metabolite, Metyrapone, Microsomal, Microsomal incubations, Microsomal metabolism, Microsomal metabolites, Microsome, Nadph, Preincubation, Present study, Prominent ions, Protein concentration, Quantitation, Radiochemical, Radiochemical detection, Radiochemical quantitation, Scintillant, Trimethylsilyl ether methyl ester, Udya.
Abstract
Abstract: An assay for the microsomal hydroxylation of lauric acid (LA), based on HPLC with flow-through radiochemical detection, has been developed. Conditions were optimized for resolution and quantitation of three microsomal metabolites of LA, one of which has not been reported previously as a metabolite of LA in mammalian microsomal incubations. These products, 12-(ω)-hydroxy-LA, 11-(ω-1)-hydroxy-LA, and a novel metabolite, 10-(ω-2)-hydroxy-LA, were isolated by HPLC and identified by gas chromatography/mass spectrometry. In the presence of NADPH, the formation of all three metabolites was linear with time and microsomal protein concentration. Hydrogen peroxide also supported the microsomal metabolism of LA, although the ratio of metabolites was substantially different than that produced by NADPH-supported microsomes. Several biochemical probes (metyrapone, α-naphthoflavone, 2-diethylaminoethyl-2,2-diphenylvalerate hydrochloride, and 10-undecynoic acid) were used to dissociate the three LA hydroxylase activities. These experiments suggest that the site-specific hydroxylation [ω-, (ω-1)-, (ω-2)-] of LA may be catalyzed by different isozymes of cytochrome P-450.
Url:
DOI: 10.1016/0003-2697(88)90093-0
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 000653
- to stream Istex, to step Curation: 000633
- to stream Istex, to step Checkpoint: 000680
- to stream Main, to step Merge: 001563
- to stream Main, to step Curation: 001540
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Determination of microsomal lauric acid hydroxylase activity by HPLC with flow-through radiochemical quantitation</title><author><name sortKey="Romano, Maria C" sort="Romano, Maria C" uniqKey="Romano M" first="Maria C." last="Romano">Maria C. Romano</name></author><author><name sortKey="Straub, Kenneth M" sort="Straub, Kenneth M" uniqKey="Straub K" first="Kenneth M." last="Straub">Kenneth M. Straub</name></author><author><name sortKey="Yodis, Lee Ann P" sort="Yodis, Lee Ann P" uniqKey="Yodis L" first="Lee Ann P." last="Yodis">Lee Ann P. Yodis</name></author><author><name sortKey="Eckardt, Regina D" sort="Eckardt, Regina D" uniqKey="Eckardt R" first="Regina D." last="Eckardt">Regina D. Eckardt</name></author><author><name sortKey="Newton, John F" sort="Newton, John F" uniqKey="Newton J" first="John F." last="Newton">John F. Newton</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:51109367A7072979601170D1BAAA5B839244E49D</idno><date when="1988" year="1988">1988</date><idno type="doi">10.1016/0003-2697(88)90093-0</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-3GL3SHG1-B/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000653</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000653</idno><idno type="wicri:Area/Istex/Curation">000633</idno><idno type="wicri:Area/Istex/Checkpoint">000680</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">000680</idno><idno type="wicri:doubleKey">0003-2697:1988:Romano M:determination:of:microsomal</idno><idno type="wicri:Area/Main/Merge">001563</idno><idno type="wicri:Area/Main/Curation">001540</idno><idno type="wicri:Area/Main/Exploration">001540</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Determination of microsomal lauric acid hydroxylase activity by HPLC with flow-through radiochemical quantitation</title><author><name sortKey="Romano, Maria C" sort="Romano, Maria C" uniqKey="Romano M" first="Maria C." last="Romano">Maria C. Romano</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Pennsylvanie</region></placeName><wicri:cityArea>Department of Drug Metabolism, Smith Kline & French Laboratories, 709 Swedeland Road, Swedeland</wicri:cityArea></affiliation></author><author><name sortKey="Straub, Kenneth M" sort="Straub, Kenneth M" uniqKey="Straub K" first="Kenneth M." last="Straub">Kenneth M. Straub</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Pennsylvanie</region></placeName><wicri:cityArea>Department of Drug Metabolism, Smith Kline & French Laboratories, 709 Swedeland Road, Swedeland</wicri:cityArea></affiliation></author><author><name sortKey="Yodis, Lee Ann P" sort="Yodis, Lee Ann P" uniqKey="Yodis L" first="Lee Ann P." last="Yodis">Lee Ann P. Yodis</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Pennsylvanie</region></placeName><wicri:cityArea>Department of Drug Metabolism, Smith Kline & French Laboratories, 709 Swedeland Road, Swedeland</wicri:cityArea></affiliation></author><author><name sortKey="Eckardt, Regina D" sort="Eckardt, Regina D" uniqKey="Eckardt R" first="Regina D." last="Eckardt">Regina D. Eckardt</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Pennsylvanie</region></placeName><wicri:cityArea>Department of Drug Metabolism, Smith Kline & French Laboratories, 709 Swedeland Road, Swedeland</wicri:cityArea></affiliation></author><author><name sortKey="Newton, John F" sort="Newton, John F" uniqKey="Newton J" first="John F." last="Newton">John F. Newton</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Pennsylvanie</region></placeName><wicri:cityArea>Department of Drug Metabolism, Smith Kline & French Laboratories, 709 Swedeland Road, Swedeland</wicri:cityArea></affiliation></author></analytic><monogr></monogr><series><title level="j">Analytical Biochemistry</title><title level="j" type="abbrev">YABIO</title><idno type="ISSN">0003-2697</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1988">1988</date><biblScope unit="volume">170</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="83">83</biblScope><biblScope unit="page" to="93">93</biblScope></imprint><idno type="ISSN">0003-2697</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0003-2697</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>HPLC, lipids</term><term>cytochrome P-450</term><term>lauric acid</term><term>metabolism</term><term>microsomes</term><term>ω-hydroxylation</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>Academic press</term><term>Assay</term><term>Biol</term><term>Chem</term><term>Control values</term><term>Cytochrome</term><term>Different isozymes</term><term>Enzyme activities</term><term>Ether</term><term>Flow cell size</term><term>Flow rate</term><term>Glucose</term><term>Hepatic</term><term>Hepatic microsomes</term><term>Hplc</term><term>Hplc effluent ratio</term><term>Hydrogen peroxide</term><term>Hydroxylase</term><term>Hydroxylase activities</term><term>Hydroxylases</term><term>Hydroxylated metabolites</term><term>Hydroxylation</term><term>Incubation</term><term>Isozymes</term><term>Kupfer</term><term>Lauric</term><term>Lauric acid</term><term>Lauric acid metabolites</term><term>Metabolism</term><term>Metabolite</term><term>Metyrapone</term><term>Microsomal</term><term>Microsomal incubations</term><term>Microsomal metabolism</term><term>Microsomal metabolites</term><term>Microsome</term><term>Nadph</term><term>Preincubation</term><term>Present study</term><term>Prominent ions</term><term>Protein concentration</term><term>Quantitation</term><term>Radiochemical</term><term>Radiochemical detection</term><term>Radiochemical quantitation</term><term>Scintillant</term><term>Trimethylsilyl ether methyl ester</term><term>Udya</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: An assay for the microsomal hydroxylation of lauric acid (LA), based on HPLC with flow-through radiochemical detection, has been developed. Conditions were optimized for resolution and quantitation of three microsomal metabolites of LA, one of which has not been reported previously as a metabolite of LA in mammalian microsomal incubations. These products, 12-(ω)-hydroxy-LA, 11-(ω-1)-hydroxy-LA, and a novel metabolite, 10-(ω-2)-hydroxy-LA, were isolated by HPLC and identified by gas chromatography/mass spectrometry. In the presence of NADPH, the formation of all three metabolites was linear with time and microsomal protein concentration. Hydrogen peroxide also supported the microsomal metabolism of LA, although the ratio of metabolites was substantially different than that produced by NADPH-supported microsomes. Several biochemical probes (metyrapone, α-naphthoflavone, 2-diethylaminoethyl-2,2-diphenylvalerate hydrochloride, and 10-undecynoic acid) were used to dissociate the three LA hydroxylase activities. These experiments suggest that the site-specific hydroxylation [ω-, (ω-1)-, (ω-2)-] of LA may be catalyzed by different isozymes of cytochrome P-450.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Pennsylvanie</li></region></list><tree><country name="États-Unis"><region name="Pennsylvanie"><name sortKey="Romano, Maria C" sort="Romano, Maria C" uniqKey="Romano M" first="Maria C." last="Romano">Maria C. Romano</name></region><name sortKey="Eckardt, Regina D" sort="Eckardt, Regina D" uniqKey="Eckardt R" first="Regina D." last="Eckardt">Regina D. Eckardt</name><name sortKey="Newton, John F" sort="Newton, John F" uniqKey="Newton J" first="John F." last="Newton">John F. Newton</name><name sortKey="Straub, Kenneth M" sort="Straub, Kenneth M" uniqKey="Straub K" first="Kenneth M." last="Straub">Kenneth M. Straub</name><name sortKey="Yodis, Lee Ann P" sort="Yodis, Lee Ann P" uniqKey="Yodis L" first="Lee Ann P." last="Yodis">Lee Ann P. Yodis</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Sante/explor/CovidV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001540 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 001540 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Sante
|area= CovidV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= ISTEX:51109367A7072979601170D1BAAA5B839244E49D
|texte= Determination of microsomal lauric acid hydroxylase activity by HPLC with flow-through radiochemical quantitation
}}
| This area was generated with Dilib version V0.6.33. |  |