Combining Amine Metabolomics and Quantitative Proteomics of Cancer Cells Using Derivatization with Isobaric Tags

Overview

Journal Anal Chem

Specialty Chemistry

Date 2014 Mar 12

PMID 24611633

Citations 20

Authors

J Patrick Murphy

Robert A Everley

Jonathan L Coloff

Steven P Gygi

Affiliations

Soon will be listed here.

Abstract

Quantitative metabolomics and proteomics technologies are powerful approaches to explore cellular metabolic regulation. Unfortunately, combining the two technologies typically requires different LC-MS setups for sensitive measurement of metabolites and peptides. One approach to enhance the analysis of certain classes of metabolites is by derivatization with various types of tags to increase ionization and chromatographic efficiency. We demonstrate here that derivatization of amine metabolites with tandem mass tags (TMT), typically used in multiplexed peptide quantitation, facilitates amino acid analysis by standard nanoflow reversed-phase LC-MS setups used for proteomics. We demonstrate that this approach offers the potential to perform experiments at the MS1-level using duplex tags or at the MS2-level using novel 10-plex reporter ion-containing isobaric tags for multiplexed amine metabolite analysis. We also demonstrate absolute quantitative measurements of amino acids conducted in parallel with multiplexed quantitative proteomics, using similar LC-MS setups to explore cellular amino acid regulation. We further show that the approach can also be used to determine intracellular metabolic labeling of amino acids from glucose carbons.

Citing Articles

Targeted and Untargeted Amine Metabolite Quantitation in Single Cells with Isobaric Multiplexing.

Heininen J, Movahedi P, Kotiaho T, Kostiainen R, Pahikkala T, Teppo J Chemistry. 2024; 30(72):e202403278.

PMID: 39422672 PMC: 11665497. DOI: 10.1002/chem.202403278.

Polarity-regulated derivatization-assisted LC-MS method for amino-containing metabolites profiling in gastric cancer.

Han J, Gong S, Bian X, Qian Y, Wang G, Li N J Pharm Anal. 2024; 13(11):1353-1364.

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Hu E, Tang T, Li Y, Li T, Zhu L, Ding R CNS Neurosci Ther. 2023; 30(3):e14231.

PMID: 37183394 PMC: 10915989. DOI: 10.1111/cns.14231.

Quantification of Serum Metabolites in Early Colorectal Adenomas Using Isobaric Labeling Mass Spectrometry.

Liu Y, Zhang H, Dove W, Wang Z, Zhu Z, Pickhardt P J Proteome Res. 2023; 22(5):1483-1491.

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