Metabolomics-PyrB enzyme assay

Description

This dataset contains targeted LC-MS/MS metabolomics data generated to assess the in vitro enzymatic activity of Escherichia coli PyrB (catalytic subunit of the aspartate carbamoyltransferase). The primary objective of this assay was to evaluate the kinetic responses of PyrB catalysis under varying concentrations of the small-molecule drug duloxetine (0–10 μM) or total RNA extract (0–70 μg/mL). Enzymatic reactions were initiated using L-aspartate (0–50 mM) and carbamoyl phosphate (48 mM) as substrates in a Tris-Acetate buffer. Following targeted quenching, metabolite separation and quantification were achieved using hydrophilic interaction liquid chromatography (HILIC) on an Atlantis™ Premier BEH Z-HILIC column. Data acquisition was performed utilizing an Agilent 1290 Infinity II UPLC system coupled to an Agilent 6470 triple quadrupole (QQQ) mass spectrometer operating in dynamic multiple reaction monitoring (MRM) mode. The dataset includes the raw mass spectrometry files and peak integration data used to quantify the specific targeted metabolites representing PyrB turnover.  arrow_drop_uphide

Institutions

• University of Cambridge

  England, Cambridge

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