دورية أكاديمية
Kinetics of dextromethorphan-O-demethylase activity and distribution of CYP2D in four commonly-used subcellular fractions of rat brain.
| العنوان: | Kinetics of dextromethorphan-O-demethylase activity and distribution of CYP2D in four commonly-used subcellular fractions of rat brain. |
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| المؤلفون: | DuBois BN; a Department of Biomedical and Pharmaceutical Sciences, School of Pharmacy , Chapman University , Irvine , CA , USA., Amirrad F; a Department of Biomedical and Pharmaceutical Sciences, School of Pharmacy , Chapman University , Irvine , CA , USA., Mehvar R; a Department of Biomedical and Pharmaceutical Sciences, School of Pharmacy , Chapman University , Irvine , CA , USA. |
| المصدر: | Xenobiotica; the fate of foreign compounds in biological systems [Xenobiotica] 2019 Oct; Vol. 49 (10), pp. 1133-1142. Date of Electronic Publication: 2019 Jan 07. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Informa Healthcare Country of Publication: England NLM ID: 1306665 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1366-5928 (Electronic) Linking ISSN: 00498254 NLM ISO Abbreviation: Xenobiotica Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: London : Informa Healthcare Original Publication: London, Taylor & Francis. |
| مواضيع طبية MeSH: | Brain/*enzymology , Cytochrome P-450 Enzyme System/*metabolism , Cytochrome P450 Family 2/*metabolism , Microsomes/*enzymology , Mitochondria/*enzymology , Oxidoreductases, O-Demethylating/*metabolism, Animals ; Brain Chemistry ; Cytochrome P-450 Enzyme System/chemistry ; Cytochrome P450 Family 2/chemistry ; Kinetics ; Male ; Oxidoreductases, O-Demethylating/chemistry ; Rats ; Rats, Sprague-Dawley |
| مستخلص: | The purpose of this study was to compare the enzymatic kinetics and distribution of cytochrome P450 2D (CYP2D) among different rat brain subcellular fractions. Rat brains were used to prepare total membrane, crude mitochondrial, purified mitochondrial, and microsomal fractions, in addition to total homogenate. Michaelis-Menten kinetics of the brain CYP2D activity was estimated based on the conversion of dextromethorphan (DXM) to dextrorphan using UPLC-MS/MS. Protein levels of CYP2D and subcellular markers were determined by Western blot. Microsomal CYP2D exhibited high affinity and low capacity, compared with the mitochondrial CYP2D that had a much lower (∼50-fold) affinity but a higher (∼six-fold) capacity. The apparent CYP2D affinity and capacity of the crude mitochondria were in between those of the microsomes and purified mitochondria. Additionally, the CYP2D activity in the whole homogenate was much higher than that in the total membranes at higher DXM concentrations. A CYP2D immune-reactive band in the brain mitochondria appeared at a lower MW but had a much higher intensity than that in the microsomes. Mitochondrial brain CYP2D has a much higher capacity than its microsomal counterpart. Additionally, brain homogenate is more representative of the overall CYP2D activity than the widely-used total membrane fraction. |
| فهرسة مساهمة: | Keywords: CYP2D; Michaelis-Menten kinetics; dextromethorphan; dextromethorphan-O-demethylase activity; drug metabolism; microsomes; mitochondria; rat brain |
| المشرفين على المادة: | 9035-51-2 (Cytochrome P-450 Enzyme System) EC 1.- (Oxidoreductases, O-Demethylating) EC 1.- (dextromethorphan O-demethylase) EC 1.14.14.1 (Cytochrome P450 Family 2) |
| تواريخ الأحداث: | Date Created: 20181106 Date Completed: 20191219 Latest Revision: 20191219 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1080/00498254.2018.1539782 |
| PMID: | 30392427 |
| قاعدة البيانات: | MEDLINE |
PDF full text from Publisher | تدمد: | 1366-5928 |
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| DOI: | 10.1080/00498254.2018.1539782 |