PMID- 30392427
OWN - NLM
STAT- MEDLINE
DCOM- 20191219
LR  - 20191219
IS  - 1366-5928 (Electronic)
IS  - 0049-8254 (Linking)
VI  - 49
IP  - 10
DP  - 2019 Oct
TI  - Kinetics of dextromethorphan-O-demethylase activity and distribution of CYP2D in 
      four commonly-used subcellular fractions of rat brain.
PG  - 1133-1142
LID - 10.1080/00498254.2018.1539782 [doi]
AB  - 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 ( approximately 50-fold) affinity but a higher 
      ( approximately 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.
FAU - DuBois, Barent N
AU  - DuBois BN
AD  - a Department of Biomedical and Pharmaceutical Sciences, School of Pharmacy , 
      Chapman University , Irvine , CA , USA.
FAU - Amirrad, Farideh
AU  - Amirrad F
AD  - a Department of Biomedical and Pharmaceutical Sciences, School of Pharmacy , 
      Chapman University , Irvine , CA , USA.
FAU - Mehvar, Reza
AU  - Mehvar R
AD  - a Department of Biomedical and Pharmaceutical Sciences, School of Pharmacy , 
      Chapman University , Irvine , CA , USA.
LA  - eng
PT  - Journal Article
DEP - 20190107
PL  - England
TA  - Xenobiotica
JT  - Xenobiotica; the fate of foreign compounds in biological systems
JID - 1306665
RN  - 9035-51-2 (Cytochrome P-450 Enzyme System)
RN  - EC 1.- (Oxidoreductases, O-Demethylating)
RN  - EC 1.- (dextromethorphan O-demethylase)
RN  - EC 1.14.14.1 (Cytochrome P450 Family 2)
SB  - IM
MH  - Animals
MH  - Brain/*enzymology
MH  - Brain Chemistry
MH  - Cytochrome P-450 Enzyme System/chemistry/*metabolism
MH  - Cytochrome P450 Family 2/chemistry/*metabolism
MH  - Kinetics
MH  - Male
MH  - Microsomes/*enzymology
MH  - Mitochondria/*enzymology
MH  - Oxidoreductases, O-Demethylating/chemistry/*metabolism
MH  - Rats
MH  - Rats, Sprague-Dawley
OTO - NOTNLM
OT  - CYP2D
OT  - Michaelis-Menten kinetics
OT  - dextromethorphan
OT  - dextromethorphan-O-demethylase activity
OT  - drug metabolism
OT  - microsomes
OT  - mitochondria
OT  - rat brain
EDAT- 2018/11/06 06:00
MHDA- 2019/12/20 06:00
CRDT- 2018/11/06 06:00
PHST- 2018/11/06 06:00 [pubmed]
PHST- 2019/12/20 06:00 [medline]
PHST- 2018/11/06 06:00 [entrez]
AID - 10.1080/00498254.2018.1539782 [doi]
PST - ppublish
SO  - Xenobiotica. 2019 Oct;49(10):1133-1142. doi: 10.1080/00498254.2018.1539782. Epub 
      2019 Jan 7.