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Profiling the Metabolism of Human Cells by Deep ¹³C Labeling

23 Pages Posted: 2 Apr 2018 Sneak Peek Status: Published

See all articles by Nina Grankvist

Nina Grankvist

Karolinska Institutet - Cardiovascular Medicine Unit

Jeramie D. Watrous

University of California, San Diego (UCSD), School of Medicine, Department of Pathology

Kim A. Lehmann

University of California, San Diego (UCSD), School of Medicine, Department of Pathology

Yaroslav Lyutvinskiy

Karolinska Institutet - Cardiovascular Medicine Unit

Mohit Jain

University of California, San Diego (UCSD), School of Medicine, Department of Pathology

Roland Nilsson

Karolinska Institutet - Cardiovascular Medicine Unit

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Abstract

The metabolism of most human cell types remains largely unexplored, in part due to lack of methods for measuring metabolic activities in an unbiased, hypothesis-free manner. Here we introduce the deep labeling method for profiling metabolic activities, based on a custom ¹³C medium in combination with high-resolution mass spectrometry. A proof-of-principle study on human cancer cells revealed hundreds of endogenous metabolites and identified a variety of metabolic activities, but also lack of central pathways synthesizing cysteine, carnitine and creatine. Protein and nucleic acids were almost exclusively de novo synthesized, while lipids were partly derived from serum fat. Branched-chain keto acids (BCKA) were formed and metabolized to a short-chain acylcarnitines, but did not enter the TCA cycle. Remarkably, BCKA could substitute for essential amino acids to support growth of these cells. We anticipate that deep labeling can be used to catalogue metabolic phenotypes across a range of cell types and conditions.

Suggested Citation

Grankvist, Nina and Watrous, Jeramie D. and Lehmann, Kim A. and Lyutvinskiy, Yaroslav and Jain, Mohit and Nilsson, Roland, Profiling the Metabolism of Human Cells by Deep ¹³C Labeling (2018). Available at SSRN: https://ssrn.com/abstract=3152002 or http://dx.doi.org/10.2139/ssrn.3152002
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Nina Grankvist (Contact Author)

Karolinska Institutet - Cardiovascular Medicine Unit

Stockholm, SE-171 77
Sweden

Jeramie D. Watrous

University of California, San Diego (UCSD), School of Medicine, Department of Pathology

9500 Gilman Drive
La Jolla, CA 92093-0636
United States

Kim A. Lehmann

University of California, San Diego (UCSD), School of Medicine, Department of Pathology

9500 Gilman Drive
La Jolla, CA 92093-0636
United States

Yaroslav Lyutvinskiy

Karolinska Institutet - Cardiovascular Medicine Unit

Stockholm, SE-171 77
Sweden

Mohit Jain

University of California, San Diego (UCSD), School of Medicine, Department of Pathology

9500 Gilman Drive
La Jolla, CA 92093-0636
United States

Roland Nilsson

Karolinska Institutet - Cardiovascular Medicine Unit ( email )

Stockholm, SE-171 77
Sweden