Tracing 15N with chemical reaction interface mass spectrometry: A demonstration using 15N-labeled glutamine and asparagine substrates in cell culture
✍ Scribed by Jozef J. Kusmierz; Fred P. Abramson
- Book ID
- 102773778
- Publisher
- John Wiley and Sons
- Year
- 1994
- Tongue
- English
- Weight
- 666 KB
- Volume
- 23
- Category
- Article
- ISSN
- 1076-5174
No coin nor oath required. For personal study only.
✦ Synopsis
This research demonstrates how the chemical reaction interface mass spectrometry (CRIMS) approach works for a study of amino acid metabolism in cell culture. ''N-selective chromatograms from both the culture medium and the cytosol of human hepatoma Hep G2 cells that were incubated in the presence of either 12 mM (a-''N)glutamine or (a-"N)asparagine have been produced. The time course of the distribution of "N among different amino acids, as well as the enrichment for each amino acid, were observed over a 144 h period. Labeled glutamine was quickly converted into glutamate. After 144 h of incubation, the total amount of "N was distributed primarily among alanine (50%), proline (28%) and glutamate (21%). The "N enrichment of alanine and proline reached 44% and 41% respectively. Asparagine was only slowly metabolized by the cells. In addition to the 82% that was retained in asparagine, the remaining "N in the media at 144 h was found primarily in alanine (So/.), glutamate (6.8%) and proline (2.2%). Their enrichments were 20%, 36% and 19% respectively. The minimum detectable amount was 17 pg of "N entering the CRI. CRIMS appears to be a powerful, facile approacb for I 'N-tracer experiments.