Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here for more information

Sign In to gain access to subscriptions and/or personal tools.
Journal of Biomolecular Screening
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
1087057109345528v1
14/9/1129    most recent
Right arrow References
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Dhayalan, A.
Right arrow Articles by Jeltsch, A.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Dhayalan, A.
Right arrow Articles by Jeltsch, A.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

A Continuous Protein Methyltransferase (G9a) Assay for Enzyme Activity Measurement and Inhibitor Screening

Arunkumar Dhayalan

Biochemistry Laboratory, School of Engineering and Science, Jacobs University Bremen, Bremen, Germany

Emilia Dimitrova

Biochemistry and Cell Biology Program, School of Engineering and Science, Jacobs University Bremen, Bremen, Germany

Philipp Rathert

Biochemistry Laboratory, School of Engineering and Science, Jacobs University Bremen, Bremen, Germany

Albert Jeltsch

Biochemistry Laboratory, School of Engineering and Science, Jacobs University Bremen, Bremen, Germany, a.jeltsch{at}jacobs-university.de

The authors describe a continuous protein methylation assay using the G9a protein lysine methyltransferase and its substrate protein WIZ (widely interspaced zinc finger motifs). The assay is based on the coupling of the biotinylated substrate protein to streptavidin-coated FlashPlates and the transfer of radioactive methyl groups from the S-adenosyl-L-methionine to the substrate. The reaction progress is monitored continuously by proximity scintillation counting. The assay is very accurate, convenient, well suited for automation, and highly reproducible with standard errors in the range of 5%. Because of few pipetting steps and continuous data readout, it is ideal for high-throughput applications such as screening of inhibitors, testing many enzyme variants, or analyzing differences in methylation rates of different substrates under various conditions. By using this new assay, the IC 50 of AdoHcy and the G9a inhibitor BIX-01294 were determined for methylation of the G9a nonhistone substrate WIZ. (Journal of Biomolecular Screening 2009:1129-1133)

Key Words: protein modifications • enzyme inhibitor • protein methyltransferase • inhibitor screening • enzyme catalysis

This version was published on October 1, 2009

Journal of Biomolecular Screening, Vol. 14, No. 9, 1129-1133 (2009)
DOI: 10.1177/1087057109345528
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?