Metabolomics-PyrB enzyme assay

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本数据集包含靶向液相色谱-串联质谱（LC-MS/MS）代谢组学数据，用于评估大肠杆菌（Escherichia coli）PyrB（天冬氨酸氨甲酰转移酶的催化亚基）的体外酶活性。本实验的首要目标是评估PyrB催化反应在不同浓度的小分子药物度洛西汀（0–10 μM）或总RNA提取物（0–70 μg/mL）条件下的动力学响应。酶促反应以L-天冬氨酸（0–50 mM）与氨甲酰磷酸（48 mM）为底物，在Tris-乙酸缓冲液中启动。经靶向淬灭后，通过Atlantis™ Premier BEH Z-HILIC色谱柱搭载的亲水相互作用液相色谱（HILIC）系统完成代谢物的分离与定量。数据采集通过安捷伦1290 Infinity II超高效液相色谱（UPLC）系统，与运行于动态多反应监测（MRM）模式的安捷伦6470三重四极杆（QQQ）质谱仪联用完成。本数据集包含用于定量表征PyrB代谢周转的特定靶向代谢物的原始质谱文件与峰积分数据。