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Analysis of nucleotide pools in bacteria using HPLC-MS in HILIC mode
E. Zborníková, Z. Knejzlík, V. Hauryliuk, L. Krásný, D. Rejman,
Jazyk angličtina Země Nizozemsko
Typ dokumentu časopisecké články
Grantová podpora
NV17-29680A
MZ0
CEP - Centrální evidence projektů
- MeSH
- Escherichia coli K12 izolace a purifikace MeSH
- hmotnostní spektrometrie metody MeSH
- koncentrace vodíkových iontů MeSH
- limita detekce MeSH
- nukleotidy chemie MeSH
- rozpouštědla chemie MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
Nucleotides, nucleosides and their derivatives are present in all cells at varying concentrations that change with the nutritional, and energetic status of the cell. Precise measurement of the concentrations of these molecules is instrumental for understanding their regulatory effects. Such measurement is challenging due to the inherent instability of these molecules and, despite many decades of research, the reported values differ widely. Here, we present a comprehensive and easy-to-use approach for determination of the intracellular concentrations of >25 target molecular species. The approach uses rapid filtration and cold acidic extraction followed by high performance liquid chromatography (HPLC) in the hydrophilic interaction liquid chromatography (HILIC) mode using zwitterionic columns coupled with UV and MS detectors. The method reliably detects and quantifies all the analytes expected to be observed in the bacterial cell and paves the way for future studies correlating their concentrations with biological effects.
Citace poskytuje Crossref.org
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- $a Zborníková, Eva $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i., Flemingovonam. 2, CZ-166 10, Prague 6, Czech Republic; Charles University, Faculty of Science, Department of Analytical Chemistry, Hlavova 8, 128 43, Prague 2, Czech Republic.
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- $a Nucleotides, nucleosides and their derivatives are present in all cells at varying concentrations that change with the nutritional, and energetic status of the cell. Precise measurement of the concentrations of these molecules is instrumental for understanding their regulatory effects. Such measurement is challenging due to the inherent instability of these molecules and, despite many decades of research, the reported values differ widely. Here, we present a comprehensive and easy-to-use approach for determination of the intracellular concentrations of >25 target molecular species. The approach uses rapid filtration and cold acidic extraction followed by high performance liquid chromatography (HPLC) in the hydrophilic interaction liquid chromatography (HILIC) mode using zwitterionic columns coupled with UV and MS detectors. The method reliably detects and quantifies all the analytes expected to be observed in the bacterial cell and paves the way for future studies correlating their concentrations with biological effects.
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- $a Knejzlík, Zdeněk $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i., Flemingovonam. 2, CZ-166 10, Prague 6, Czech Republic.
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- $a Hauryliuk, Vasili $u Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87, Umeå, Sweden; Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Building 6K and 6L, University Hospital Area, 90187, Umeå, Sweden; University of Tartu, Institute of Technology, 50411, Tartu, Estonia.
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- $a Krásný, Libor $u Institute of Microbiology, Czech Academy of Sciences v.v.i., Vídeňská 1083, 142 20, Prague 4, Czech Republic.
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- $a Rejman, Dominik $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i., Flemingovonam. 2, CZ-166 10, Prague 6, Czech Republic. Electronic address: rejman@uochb.cas.cz.
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