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
Molecular Pharmacology
  • 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
Molecular Pharmacology

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 molpharm on Facebook
  • Follow molpharm on Twitter
  • Follow molpharm on LinkedIn
OtherAccelerated Communication

An Active and Water-Soluble Truncation Mutant of the Human UDP-Glucuronosyltransferase 1A9

Mika Kurkela, Saila Mörsky, Jouni Hirvonen, Risto Kostiainen and Moshe Finel
Molecular Pharmacology April 2004, 65 (4) 826-831; DOI: https://doi.org/10.1124/mol.65.4.826
Mika Kurkela
Viikki Drug Discovery Technology Center (M.K., S.M., J.H., R.K., M.F.), Faculty of Pharmacy (J.H., R.K.), University of Helsinki, Helsinki, Finland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Saila Mörsky
Viikki Drug Discovery Technology Center (M.K., S.M., J.H., R.K., M.F.), Faculty of Pharmacy (J.H., R.K.), University of Helsinki, Helsinki, Finland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jouni Hirvonen
Viikki Drug Discovery Technology Center (M.K., S.M., J.H., R.K., M.F.), Faculty of Pharmacy (J.H., R.K.), University of Helsinki, Helsinki, Finland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Risto Kostiainen
Viikki Drug Discovery Technology Center (M.K., S.M., J.H., R.K., M.F.), Faculty of Pharmacy (J.H., R.K.), University of Helsinki, Helsinki, Finland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Moshe Finel
Viikki Drug Discovery Technology Center (M.K., S.M., J.H., R.K., M.F.), Faculty of Pharmacy (J.H., R.K.), University of Helsinki, Helsinki, Finland
  • 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

The UDP-glucuronosyltransferases (UGTs) are integral membrane proteins, and previous attempts to generate a water-soluble UGT by removing the single trans-membrane helix yielded inactive and membrane-bound proteins. We have now replaced the 45 C-terminal amino acids of the human UGT1A9, including its trans-membrane helix, with a fusion peptide ending with six His residues. Detergent-free extraction of insect cells expressing this mutant, UGT1A9Sol, released scopoletin glucuronidation activity into the supernatant, and subsequent ultracentrifugation did not sediment that activity. UGT1A9Sol was purified by immobilized metal affinity chromatography (IMAC) in the absence of detergents throughout the entire process. The IMAC purification increased somewhat the apparent Km of UGT1A9 toward scopoletin and rendered the enzyme sensitive to freezing. The activity of UGT1A9Sol in the cell extract was partly inhibited by Triton X-100, irrespective of the presence or absence of phospholipids. UGT1A9Sol exhibited a relatively high rate of scopoletin glucuronidation, whereas its activity toward 1-naphthol, entacapone, umbelliferone, and 4-nitrophenol was much lower. The kinetics and substrate specificity of UGT1A9Sol resembled the detergent-suspended full-length UGT1A9 rather than the membrane-bound UGT1A9. The apparent Km value of UGT1A9Sol for scopoletin was similar to that of the full-length UGT1A9 in the presence of detergent, but much higher than the respective value in the membrane-bound enzyme. The results suggest that either the detergent binding to the trans-membrane helix within the full-length UGT1A9, or the removal of this helix by gene manipulation, affect the interaction of the enzyme with its aglycone substrate in a similar manner.

  • The American Society for Pharmacology and Experimental Therapeutics
View Full Text

MolPharm 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

Molecular Pharmacology: 65 (4)
Molecular Pharmacology
Vol. 65, Issue 4
1 Apr 2004
  • Table of Contents
  • About the Cover
  • Index by author
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Molecular Pharmacology 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.
An Active and Water-Soluble Truncation Mutant of the Human UDP-Glucuronosyltransferase 1A9
(Your Name) has forwarded a page to you from Molecular Pharmacology
(Your Name) thought you would be interested in this article in Molecular Pharmacology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
OtherAccelerated Communication

An Active and Water-Soluble Truncation Mutant of the Human UDP-Glucuronosyltransferase 1A9

Mika Kurkela, Saila Mörsky, Jouni Hirvonen, Risto Kostiainen and Moshe Finel
Molecular Pharmacology April 1, 2004, 65 (4) 826-831; DOI: https://doi.org/10.1124/mol.65.4.826

Citation Manager Formats

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

Share
OtherAccelerated Communication

An Active and Water-Soluble Truncation Mutant of the Human UDP-Glucuronosyltransferase 1A9

Mika Kurkela, Saila Mörsky, Jouni Hirvonen, Risto Kostiainen and Moshe Finel
Molecular Pharmacology April 1, 2004, 65 (4) 826-831; DOI: https://doi.org/10.1124/mol.65.4.826
del.icio.us logo Digg logo Reddit logo Twitter logo Facebook logo Google 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

  • GABAA Receptor Desensitization by Low GABA
  • Structure of the Diltiazem Receptor Site on Calcium Channels
  • 5-HT and Sleep
Show more Accelerated Communication

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

ASPET's Other Journals

  • Drug Metabolism and Disposition
  • Journal of Pharmacology and Experimental Therapeutics
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-0111 (Online)

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