A High Throughput Assay to Identify Small Molecule Modulators of Prostatic Acid Phosphatase
Rylan S Larsen1, Mark J Zylka1, John E Scott*, 2
1 Department of Cell and Molecular Physiology, UNC Neuroscience Center, University of North Carolina at Chapel Hill, CB#7545, Chapel Hill, NC 27599, USA
2 Department of Pharmaceutical Sciences, Biomanufacturing Research Institute and Technology Enterprise, North Carolina Central University, 1801 Fayetteville Street, Durham, NC 27707, USA
Abstract
Prostatic acid phosphatase (PAP) is expressed in nociceptive neurons and functions as an ectonucleotidase. Injection of the secretory isoform of PAP has potent antinociceptive effects in mouse models of chronic pain. These data suggested that a small molecule activator of PAP may have utility as a novel therapeutic for chronic pain, while inhibitors could be used to acutely inhibit PAP in vitro and in vivo. To identify small molecule modulators of PAP activity, we validated a high throughput, fluorescence-based biochemical assay and then used this assay to screen a compound library. We decreased the frequency of false positive activators by subtracting compound fluorescence from the final assay fluorescence. This approach significantly reduced the number of false positive activators found in the screen. While no activators were confirmed, seven novel inhibitors of PAP were identified. Our results suggest this high throughput assay could be used to identify small molecule modulators of PAP activity.
Article Information
Article History:
Received Date: 17/3/2009
Revision Received Date: 30/4/2009
Acceptance Date: 2/5/2009
Electronic publication date: 16/6/2009
Collection year: 2009
© Larsen et al.; Licensee Bentham Open.
open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution License (
http://creativecommons.org/licenses/by/2.5/), which
permits unrestrictive use, distribution, and reproduction in any medium, provided the original work is properly cited.
* Address correspondence to this author at the Department of Pharmaceutical Sciences, Biomanufacturing Research Institute and Technology Enterprise, North Carolina Central University, 1801 Fayetteville Street, Durham, NC 27707, USA; E-mail: jscott@nccu.edu
Open Peer Review Details |
Manuscript submitted on 17-3-2009 |
Original Manuscript |
A High Throughput Assay to Identify Small Molecule Modulators of Prostatic Acid Phosphatase |