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Myotube Protein Content Associates With Intracellular L-Glutamine Levels

Myotube Protein Content Associates With Intracellular L-Glutamine Levels

Diogo Antonio Alves de Vasconcelos, Pieter Giesbertz, Gilson Masahiro Murata, Diego Ribeiro de Souza, Jarlei Fiamoncini, Daniella Duque-Guimaraes, Carol Góis Leandro, Sandro Massao Hirabara, Hannelore Daniel, etc.

Cell Physiol Biochem. 2019;53(1):200-214.

PMID: 31287628

Abstract:

Background/aims:
Skeletal mass loss is reported in several catabolic conditions and it has been associated with a reduced intracellular L-glutamine content. We investigated the association of intracellular L-glutamine concentration with the protein content in skeletal muscle cells.
Methods:
We cultivated C2C12 myotubes in the absence or presence of 2 (reference condition), 8 or 16 mM L-glutamine for 48 hours, and the variations in the contents of amino acids and proteins measured. We used an inhibitor of L-glutamine synthesis (L-methionine sulfoximine - MSO) to promote a further reduction in intracellular L-glutamine levels. Amino acids contents in cells and media were measured using LC-MS/MS. We measured changes in phosphorylated Akt, RP-S6, and 4E-BP1contents in the absence or presence of insulin by western blotting.
Results:
Reduced intracellular L-glutamine concentration was associated with decreased protein content and increased protein breakdown. Low intracellular glutamine levels were also associated with decreased p-Akt contents in the presence of insulin. A further decrease in intracellular L-glutamine caused by glutamine synthetase inhibitor reduced protein content and levels of amino acids generated from glutamine metabolism and increased bAib still further. Cells exposed to high medium glutamine levels did not have any change in protein content but exhibited increased contents of the amino acids derived from L-glutamine metabolism.
Conclusion:
Intracellular L-glutamine levels per se play a role in the control of protein content in skeletal muscle myotubes.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP15985394	L-Methionine sulfoximine		15985-39-4	Price
AP80143573	L-Glutamine-(amine-15N)		80143-57-3	Price

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