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Hindi Research Article
A Highly Adaptable Method for Quantification of Peptidyl-tRNA Hydrolase Activity
W. Blake Holloway, Hana McFeeters, Adam M Powell, Gnana S Nidadavolu and Robert L McFeeters*
Department of Chemistry, University of Alabama in Huntsville, Huntsville, AL, USA
- *Corresponding Author:
- Robert L McFeeters
Department of Chemistry, University of Alabama in Huntsville
Huntsville, AL, USA
Tel: 256-824-6023
Fax: 256-824-6349
E-mail: robert.mcfeeters@uah.edu
Received date: March 04, 2015; Accepted date: April 28, 2015; Published date: May 06, 2015
Citation: Holloway WB, McFeeters H, Powell AM, Nidadavolu GS, McFeeters RL (2015) A Highly Adaptable Method for Quantification of Peptidyl-tRNA Hydrolase Activity. J Anal Bioanal Tech 6:244. doi: 10.4172/2155-9872.1000244
Copyright: © 2015 Holloway WB, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
The emerging importance of Peptidyl-tRNA hydrolase (Pth) enzymes necessitates the need for a widely applicable functional assay to further studies of this important enzyme family. Previously reported methods for monitoring Pth function suffer from limitations of cost, time, substrate availability, and application compatibility. Herein we present a new method for the rapid and precise characterization of Pth activity. The method is applicable for use with specific or bulk peptidyl-tRNA, any Pth enzyme, and a range of reaction conditions including solvent additives. The method also allows for semi-automated quantitative assessment of peptidyl-tRNA cleavage. No specialized equipment, harmful reagents, or time-consuming techniques are required. We use the new method to characterize Pth activity, determine enzyme kinetic parameters, screen for inhibitors, and determine inhibitory parameters.
