Document Detail


Characterization of CYP2A6 involved in 3'-hydroxylation of cotinine in human liver microsomes.
MedLine Citation:
PMID:  8627511     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Nicotine is primarily metabolized to cotinine, and cotinine is further metabolized to trans-3'-hydroxycotinine in human liver, which is a major metabolite of nicotine in humans. We studied the formation of trans-3'-hydroxycotinine from cotinine in human liver microsomes. trans-3'-Hydroxycotinine formation demonstrated single enzyme Michaelis-Menten kinetics (Km, 234.5 +/- 26.8 MicroM; Vmax, 37.2 +/- 2.4 pmol/min/mg protein). Significant correlation (r = .967, P < .001) between cotinine 3'-hydroxylase activities at low (50 microM) and high (1 microM) cotinine concentrations in 20 human liver microsomes suggested the contribution of a single enzyme to cotinine 3'-hydroxylation. The cotinine 3'-hydroxylase activity correlated significantly with immunoreactive cytochrome P450 (CYP)2A6 contents (r = .756, P < .01) and coumarin 7-hydroxylase activity (r = .887, P < .001). The cotinine 3'-hydroxylase activity was inhibited by coumarin, alpha-naphthoflavone, chlorzoxazone and anti-rat CYP2A1 antibodies. Microsomes of B-lymphoblastoid cells expressing human CYP2A6 exhibited cotinine 3'-hydroxylase activity. The Km value of the expressed CYP2A6 (264.7 microM) was almost identical to that of human liver microsomes. In conclusion, cotinine 3'-hydroxylation appears to be catalyzed solely by CYP2A6 in humans. Cotinine is a candidate for a new substrate for CYP2A6 in humans.
Authors:
M Nakajima; T Yamamoto; K Nunoya; T Yokoi; K Nagashima; K Inoue; Y Funae; N Shimada; T Kamataki; Y Kuroiwa
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of pharmacology and experimental therapeutics     Volume:  277     ISSN:  0022-3565     ISO Abbreviation:  J. Pharmacol. Exp. Ther.     Publication Date:  1996 May 
Date Detail:
Created Date:  1996-06-27     Completed Date:  1996-06-27     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0376362     Medline TA:  J Pharmacol Exp Ther     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1010-5     Citation Subset:  IM    
Affiliation:
Department of Clinical Pharmacy, School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
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MeSH Terms
Descriptor/Qualifier:
Aryl Hydrocarbon Hydroxylases*
Cotinine / metabolism*
Cytochrome P-450 Enzyme System / antagonists & inhibitors,  physiology*
Humans
Hydroxylation
Isoenzymes / physiology
Microsomes, Liver / metabolism*
Mixed Function Oxygenases / antagonists & inhibitors,  physiology*
Chemical
Reg. No./Substance:
0/Isoenzymes; 486-56-6/Cotinine; 9035-51-2/Cytochrome P-450 Enzyme System; EC 1.-/Mixed Function Oxygenases; EC 1.14.14.1/Aryl Hydrocarbon Hydroxylases; EC 1.14.14.1/coumarin 7-hydroxylase

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