Please use this identifier to cite or link to this item:
http://hdl.handle.net/11375/14369
Full metadata record
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Bharat, Amrita | en_US |
dc.contributor.author | Blanchard, Jan E. | en_US |
dc.contributor.author | Brown, Eric | en_US |
dc.date.accessioned | 2014-06-18T17:32:47Z | - |
dc.date.available | 2014-06-18T17:32:47Z | - |
dc.date.created | 2013-07-23 | en_US |
dc.date.issued | 2013 | en_US |
dc.identifier.other | bio_chemistry_coll/1 | en_US |
dc.identifier.other | 1000 | en_US |
dc.identifier.other | 4338415 | en_US |
dc.identifier.uri | http://hdl.handle.net/11375/14369 | - |
dc.description | <p>Published in the Journal of Biomolecular Screening, doi: 10.1177/1087057113486001</p> | en_US |
dc.description.abstract | <p>The synthesis of ribosomes is an essential process, which is aided by a variety of transacting factors in bacteria. Among these is a group of GTPases essential for bacterial viability and emerging as promising targets for new antibacterial agents. Herein, we describe a robust high-throughput screening process for inhibitors of one such GTPase, the Escherichia coli EngA protein. The primary screen employed an assay of phosphate production in 384-well density.<br />Reaction conditions were chosen to maximize sensitivity for the discovery of competitive inhibitors while maintaining a strong signal amplitude and low noise. In a pilot screen of 31,800 chemical compounds, 44 active compounds were identified. Further, we describe the elimination of non-specific inhibitors that were detergent-sensitive or reactive as well as those that interfered with the high-throughput phosphate assay. Four inhibitors survived these common counterscreens for non-specificity but these chemicals were also inhibitors of the unrelated enzyme dihydrofolate reductase, suggesting that they too were promiscuously active. The highthroughput screen of the EngA protein described here provides a meticulous pilot study in the search for specific inhibitors of GTPases involved in ribosome biogenesis.</p> | en_US |
dc.subject | EngA | en_US |
dc.subject | GTPase | en_US |
dc.subject | ribosome biogenesis | en_US |
dc.subject | enzyme screen | en_US |
dc.subject | Medical Biochemistry | en_US |
dc.subject | Medical Sciences | en_US |
dc.subject | Medical Biochemistry | en_US |
dc.title | A high-throughput screen of the GTPase activity of Escherichia coli EngA to find an inhibitor of bacterial ribosome biogenesis | en_US |
dc.type | article | en_US |
Appears in Collections: | Biochemistry & Biomedical Sciences Publications |
Files in This Item:
File | Size | Format | |
---|---|---|---|
fulltext.pdf | 1.44 MB | Adobe PDF | View/Open |
Items in MacSphere are protected by copyright, with all rights reserved, unless otherwise indicated.