Please use this identifier to cite or link to this item: http://hdl.handle.net/2381/28323
Title: Optimization of a direct spectrophotometric method to investigate the kinetics and inhibition of sialidases.
Authors: Hayre, J.K.
Xu, G.
Borgianni, L.
Taylor, G.L.
Andrew, Peter W.
Docquier, J.D.
Oggioni, M.R.
First Published: 2-Oct-2012
Publisher: BioMed Central Ltd
Citation: BMC Biochemistry, 2012, 13:19
Abstract: Background: Streptococcus pneumoniae expresses three distinct sialidases, NanA, NanB, and NanC, that are believed to be key virulence factors and thus, potential important drug targets. We previously reported that the three enzymes release different products from sialosides, but could share a common catalytic mechanism before the final step of product formation. However, the kinetic investigations of the three sialidases have not been systematically done thus far, due to the lack of an easy and steady measurement of sialidase reaction rate. Results: In this work, we present further kinetic characterization of pneumococcal sialidases by using a direct spectrophotometric method with the chromogenic substrate p-nitrophenyl-N-acetylneuraminic acid (p-NP-Neu5Ac). Using our assay, the measured kinetic parameters of the three purified pneumococcal sialidase, NanA, NanB and NanC, were obtained and were in perfect agreement with the previously published data. The major advantage of this alternative method resides in the direct measurement of the released product, allowing to readily determine of initial reaction rates and record complete hydrolysis time courses. Conclusion: We developed an accurate, fast and sensitive spectrophotometric method to investigate the kinetics of sialidase-catalyzed reactions. This fast, sensitive, inexpensive and accurate method could benefit the study of the kinetics and inhibition of sialidases in general.
DOI Link: 10.1186/1471-2091-13-19
eISSN: 1471-2091
Links: http://www.biomedcentral.com/1471-2091/13/19
http://hdl.handle.net/2381/28323
Version: Publisher Version
Status: Peer-reviewed
Type: Journal Article
Rights: Copyright © 2012 Hayre et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Description: PMCID: PMC3483245
Appears in Collections:Published Articles, Dept. of Infection, Immunity and Inflammation

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