Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Submit
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET

User menu

  • My alerts
  • Log in
  • My Cart

Search

  • Advanced search
Drug Metabolism & Disposition
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET
  • My alerts
  • Log in
  • My Cart
Drug Metabolism & Disposition

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Submit
  • Visit dmd on Facebook
  • Follow dmd on Twitter
  • Follow ASPET on LinkedIn
Research ArticleArticle

mRNA Distribution and Heterologous Expression of Orphan Cytochrome P450 20A1

Katarina Stark, Zhong-Liu Wu, Cheryl J. Bartleson and F. Peter Guengerich
Drug Metabolism and Disposition September 2008, 36 (9) 1930-1937; DOI: https://doi.org/10.1124/dmd.108.022020
Katarina Stark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Zhong-Liu Wu
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Cheryl J. Bartleson
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
F. Peter Guengerich
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF
Loading

Abstract

Cytochrome P450 (P450) 20A1 is one of the so-called “orphan” P450s without assigned biological function. mRNA expression was detected in human liver, and extrahepatic expression was noted in several human brain regions, including substantia nigra, hippocampus, and amygdala, using conventional polymerase chain reaction and RNA dot blot analysis. Adult human liver contained 3-fold higher overall mRNA levels than whole brain, although specific regions (i.e., hippocampus and substantia nigra) exhibited higher mRNA expression levels than liver. Orthologous full-length and truncated transcripts of P450 20A1 were transcribed and sequenced from rat liver, heart, and brain. In rat, the concentrations of full-length transcripts were 3- to 4-fold higher in brain and heart than in liver. In situ hybridization of rat whole brain sections showed an mRNA expression pattern similar to that observed for human P450 20A1, indicating expression in substantia nigra, hippocampus, and amygdala. A number of N-terminal modifications of the codon-optimized human P450 20A1 sequence were prepared and expressed in Escherichia coli, and two of the truncated derivatives showed characteristic P450 spectra (200–280 nmol of P450/l). Although the recombinant enzyme system oxidized NADPH, no catalytic activity was observed with the heterologously expressed protein when a number of potential steroids and biogenic amines were surveyed as potential substrates. The function of P450 20A1 remains unknown; however, the sites of mRNA expression in human brain and the conservation among species may suggest possible neurophysiological function.

Footnotes

  • ↵4 In situ hybridization of human tissues was attempted; however, no conclusions could be drawn from the limited number of analyses due to lack of material.

  • This work was supported in part by the by Henning and Johan Trone Holst stiftelse (K.S.), Svenska Läkaresällskapet och Apotekarsocietéten (K.S.), and U.S. Public Health Service Grants R37 CA090426, T32 ES007028, and P30 ES000267 (F.P.G.).

  • Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.

  • doi:10.1124/dmd.108.022020.

  • ABBREVIATIONS: P450, cytochrome P450 [also termed “heme thiolate P450” (Palmer and Reedijk, 1992); CYP indicates the gene in question]; bp, base pair(s); NCBI, National Center for Biotechnology Information; SSC, 15 mM sodium citrate buffer (pH 7.0) containing 150 mM NaCl); RT, reverse transcriptase; PCR, polymerase chain reaction; GAPDH, glycerylaldehyde 3-phosphate dehydrogenase; qPCR, quantitative polymerase chain reaction; DIG, digoxigenin; TBS, Tris-buffered saline [0.10 M Tris-HCl (pH 7.5) containing 0.15 M NaCl]; UPLC, ultra-performance liquid chromatography; MS, mass spectrometry; MS/MS, tandem mass spectrometry.

  • ↵ Embedded Image The online version of this article (available at http://dmd.aspetjournals.org) contains supplemental material.

  • ↵1 Current affiliation: Experimental Asthma and Allergy Research, National Institute of Environmental Medicine, Karolinska Institute, Stockholm, Sweden.

  • ↵2 Current affiliation: Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, People's Republic of China.

  • ↵3 Current affiliation: CellzDirect Inc., Invitrogen Corporation, Austin, TX.

    • Received April 23, 2008.
    • Accepted June 4, 2008.
  • The American Society for Pharmacology and Experimental Therapeutics
View Full Text

 

DMD articles become freely available 12 months after publication, and remain freely available for 5 years. 

Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page. 

 

  • Click here for information on institutional subscriptions.
  • Click here for information on individual ASPET membership.

 

Log in using your username and password

Forgot your user name or password?

Purchase access

You may purchase access to this article. This will require you to create an account if you don't already have one.
PreviousNext
Back to top

In this issue

Drug Metabolism and Disposition: 36 (9)
Drug Metabolism and Disposition
Vol. 36, Issue 9
1 Sep 2008
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover
  • Index by author
  • Back Matter (PDF)
  • Editorial Board (PDF)
  • Front Matter (PDF)
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Drug Metabolism & Disposition article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
mRNA Distribution and Heterologous Expression of Orphan Cytochrome P450 20A1
(Your Name) has forwarded a page to you from Drug Metabolism & Disposition
(Your Name) thought you would be interested in this article in Drug Metabolism & Disposition.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Research ArticleArticle

mRNA Distribution and Heterologous Expression of Orphan Cytochrome P450 20A1

Katarina Stark, Zhong-Liu Wu, Cheryl J. Bartleson and F. Peter Guengerich
Drug Metabolism and Disposition September 1, 2008, 36 (9) 1930-1937; DOI: https://doi.org/10.1124/dmd.108.022020

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero

Share
Research ArticleArticle

mRNA Distribution and Heterologous Expression of Orphan Cytochrome P450 20A1

Katarina Stark, Zhong-Liu Wu, Cheryl J. Bartleson and F. Peter Guengerich
Drug Metabolism and Disposition September 1, 2008, 36 (9) 1930-1937; DOI: https://doi.org/10.1124/dmd.108.022020
Reddit logo Twitter logo Facebook logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Materials and Methods
    • Results
    • Discussion
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • A PBPK model for CBD in adults and children
  • Olanzapine Glucuronidation in Humanized Mice
  • rs2242480 Regulates the Expression of CYP3A4 and CYP3A5
Show more Articles

Similar Articles

Advertisement
  • Home
  • Alerts
Facebook   Twitter   LinkedIn   RSS

Navigate

  • Current Issue
  • Fast Forward by date
  • Fast Forward by section
  • Latest Articles
  • Archive
  • Search for Articles
  • Feedback
  • ASPET

More Information

  • About DMD
  • Editorial Board
  • Instructions to Authors
  • Submit a Manuscript
  • Customized Alerts
  • RSS Feeds
  • Subscriptions
  • Permissions
  • Terms & Conditions of Use

ASPET's Other Journals

  • Journal of Pharmacology and Experimental Therapeutics
  • Molecular Pharmacology
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-009X (Online)

Copyright © 2023 by the American Society for Pharmacology and Experimental Therapeutics