alexa A coupled assay measuring Mycobacterium tuberculosis antigen 85C enzymatic activity.


Medicinal chemistry

Author(s): Boucau J, Sanki AK, Voss BJ, Sucheck SJ, Ronning DR

Abstract Share this page

Abstract The prevalence of drug-resistant strains of Mycobacterium tuberculosis (M. tb) emphasizes the need for new antitubercular drugs. An essential component of the drug discovery process is the development of tools to rapidly screen potential drug libraries against important biological targets. Similarly to well-documented M. tb targets, the antigen 85 (Ag85) enzymes are involved in the maintenance of the mycobacterial cell wall. The products synthesized by these mycolyltransferases are the cell wall components most responsible for the reduced permeability of drugs into the bacterial cell, thereby linking Ag85 activity directly with drug resistance. This article presents the development of a high-throughput colorimetric assay suitable for direct monitoring of the enzymatic activity. The assay uses a synthetic substrate containing three chemical moieties: an octanoyl fatty acid, beta-D-glucose, and p-nitrophenyl. In the context of the assay, Ag85 catalyzes the removal of the fatty acid and releases p-nitrophenyl-beta-D-glucoside. The glucoside is hydrolyzed by beta-glucosidase to release the p-nitrophenolate chromophore. With this assay, the K(M) and k(cat) values of Ag85C were determined to be 0.047 +/- 0.008 mM and 0.062 s(-1), respectively. In addition, the assay exhibits a Z' value of 0.81 +/- 0.06, indicating its suitability for high-throughput screening applications and drug development. This article was published in Anal Biochem and referenced in Medicinal chemistry

Relevant Expert PPTs

Relevant Speaker PPTs

Recommended Conferences

Peer Reviewed Journals
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version