期刊
ANALYTICAL CHEMISTRY
卷 87, 期 4, 页码 2129-2136出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac503210n
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资金
- Auburn University Intramural Grants Program (AU-IGP) grant through the Office of the Vice President for Research (OVPR)
- Secretaria Nacional de Ciencia y Tecnologia (SENACYT)
- Instituto para la Formacion de Recursos Humanos (IFARHU) of the Panamanian government
- Auburn University Cellular and Molecular Biosciences Undergraduate Summer Research Fellowship
A simple and reliable liquid chromatographymass spectrometry (LC-MS) assay has been developed and validated for the kinetic characterization and evaluation of inhibitors of shikimate kinase from Mycobacterium tuberculosis (MtSK), a potential target for the development of novel antitubercular drugs. This assay is based on the direct determination of the reaction product shikimate-3-phosphate (S3P) using electrospray ionization (ESI) and a quadrupole time-of-flight (Q-TOF) detector. A comparative analysis of the kinetic parameters of MtSK obtained by the LC-MS assay with those obtained by a conventional UV-assay was performed. Kinetic parameters determined by LC-MS were in excellent agreement with those obtained from the UV assay, demonstrating the accuracy, and reliability of this method. The validated assay was successfully applied to the kinetic characterization of a known inhibitor of shikimate kinase; inhibition constants and mode of inhibition were accurately delineated with LC-MS.
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