TY - JOUR
T1 - MALDI-MS-based high-throughput metabolite analysis for intracellular metabolic dynamics
AU - Yukihira, Daichi
AU - Miura, Daisuke
AU - Saito, Kazunori
AU - Takahashi, Katsutoshi
AU - Wariishi, Hiroyuki
PY - 2010/5/15
Y1 - 2010/5/15
N2 - In the present study, a high-throughput analytical method for intracellular metabolites using MALDI-MS has been developed. As an analytical tool, the quantitative performance and dynamic range of MALDI-TOF-MS was confirmed to be suitable for characterizing the trends of intracellular metabolism. The technique was tested by investigating the intracellular metabolism of Escherichia coli by analyzing whole cell samples taken consecutively before and after a perturbation of the environmental carbon source. As the result, dramatic changes of metabolite concentrations responding to the perturbation were observed. The whole analysis process (i.e., sample preparation and MALDI-MS analysis for 24 time points in triplicate) was completed within 4 hours. MALDI-FTICR-MS was used to identify the elemental compositions of detected metabolites to support the reliability of the MALDI-MS-based analysis. The MALDI-MS-based analytical method developed herein should be suitable for high-throughput analysis of dynamic intracellular metabolism events.
AB - In the present study, a high-throughput analytical method for intracellular metabolites using MALDI-MS has been developed. As an analytical tool, the quantitative performance and dynamic range of MALDI-TOF-MS was confirmed to be suitable for characterizing the trends of intracellular metabolism. The technique was tested by investigating the intracellular metabolism of Escherichia coli by analyzing whole cell samples taken consecutively before and after a perturbation of the environmental carbon source. As the result, dramatic changes of metabolite concentrations responding to the perturbation were observed. The whole analysis process (i.e., sample preparation and MALDI-MS analysis for 24 time points in triplicate) was completed within 4 hours. MALDI-FTICR-MS was used to identify the elemental compositions of detected metabolites to support the reliability of the MALDI-MS-based analysis. The MALDI-MS-based analytical method developed herein should be suitable for high-throughput analysis of dynamic intracellular metabolism events.
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U2 - 10.1021/ac100024w
DO - 10.1021/ac100024w
M3 - Article
C2 - 20408595
AN - SCOPUS:77952476087
SN - 0003-2700
VL - 82
SP - 4278
EP - 4282
JO - Analytical chemistry
JF - Analytical chemistry
IS - 10
ER -