HPLC determination of α-ketoglutaramate [5-amino-2,5-dioxopentanoate] in biological samples

Yevgeniya I Shurubor; Arthur J L Cooper; Elena P Isakova; Yulia I Deryabina; M Flint Beal; Boris F Krasnikov

doi:10.1016/j.ab.2015.11.003

HPLC determination of α-ketoglutaramate [5-amino-2,5-dioxopentanoate] in biological samples

Anal Biochem. 2016 Feb 1:494:52-4. doi: 10.1016/j.ab.2015.11.003. Epub 2015 Nov 11.

Authors

Yevgeniya I Shurubor¹, Arthur J L Cooper², Elena P Isakova³, Yulia I Deryabina³, M Flint Beal⁴, Boris F Krasnikov⁵

Affiliations

¹ The Brain and Mind Research Institute, Weill Cornell Medical College, Department of Neurology, New York, NY 10021, USA; Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, 33, bld. 2 Leninsky Ave., Moscow 119071, Russia.
² Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, USA. Electronic address: arthur_cooper@nymc.edu.
³ Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, 33, bld. 2 Leninsky Ave., Moscow 119071, Russia.
⁴ The Brain and Mind Research Institute, Weill Cornell Medical College, Department of Neurology, New York, NY 10021, USA.
⁵ Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, 33, bld. 2 Leninsky Ave., Moscow 119071, Russia; Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, USA.

PMID: 26576832
DOI: 10.1016/j.ab.2015.11.003

Abstract

α-Ketoglutaramate is an important glutamine metabolite in mammals, plants, and many bacteria. It is also a nicotine metabolite in certain bacteria. Previously published methods for the determination of α-ketoglutaramate in biological samples have considerable drawbacks. Here, we describe a relatively simple high-performance liquid chromatography (HPLC)-based method for measurement of α-ketoglutaramate in plasma and deproteinized tissues that overcomes these drawbacks. Concentrations of α-ketoglutaramate in normal rat liver, kidney, brain, and plasma were found to be approximately 216, 13, 6, and 19 μM, respectively. The HPLC method should be useful for studying the role of α-ketoglutaramate in eukaryotic glutamine metabolism and in bacterial nicotine metabolism.

Keywords: Glutaminase II pathway; α-Ketoglutaramate; ω-Amidase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Brain / metabolism
Chemistry Techniques, Analytical / methods*
Chromatography, High Pressure Liquid*
Ketoglutaric Acids / analysis*
Ketoglutaric Acids / blood
Kidney / metabolism
Liver / metabolism
Rats

Substances

Ketoglutaric Acids
alpha-ketoglutaramate