Glutamine transport in human skeletal muscle.
Sarcolemmal vesicles isolated from human skeletal muscle obtained at surgery showed approximately 14-fold enrichment of sarcolemmal marker enzymes 5'-nucleotidase and K-stimulated phosphatase. [3H]glutamine transport in these vesicles was stereospecific, largely Na dependent, and tolerated Li-for-Na substitution. Glutamine transport was stimulated by an inside negative membrane potential, and 25 mM glutamine stimulated 22Na (0.1 mM) uptake into vesicles by 50%, indicating rheogenic cotransport of Na and glutamine. Alanine transport was Na dependent but did not tolerate Li-for-Na substitution. Transport of L-[3H]glutamine was inhibited by 35-65% with a 20-fold excess of glutamine, asparagine, and alanine; cysteine, alpha-(methylamino)isobutyrate, and 2-amino-2-norborane carboxylic acid had smaller inhibitory effects, although cysteine had an unusually large inhibitory effect on glutamine transport at 1,000-fold excess compared with most other amino acids. Glutamine transport showed sensitivity to pH values < 7.0. Glutamine transport consisted of a Na-dependent and a Na-independent component, both of which appeared saturable. The kinetic characteristics of the Na-dependent component were different in different types of muscles, with half-maximal concentrations (mM) varying from 1.6 +/- 0.4 (tibialis anterior) to 0.56 +/- 0.0.2 (gluteus maximus) and maximal velocity (pmol.mg protein-1.s-1) of 1.3 +/- 0.27 to 5 +/- 1.25 in the same muscles. The results demonstrate both marked similarities and important differences between the principal glutamine transporter in human skeletal muscle and the known system Nm transporter in rat skeletal muscle.
Ahmed
A
A
Department of Anatomy and Physiology, University of Dundee, Scotland.
Maxwell
D L
DL
Taylor
P M
PM
Rennie
M J
MJ
D001216:Asparagine / Q000378:metabolism; D001692:Biological Transport; D003470:Culture Media; D005973:Glutamine / Q000378:metabolism* / Q000494:pharmacology; D006639:Histidine / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007700:Kinetics; D008564:Membrane Potentials; D009124:Muscle Proteins / Q000378:metabolism; D009132:Muscles / Q000378:metabolism* / Q000502:physiology; D009994:Osmolar Concentration; D012508:Sarcolemma / Q000378:metabolism / Q000502:physiology; D012964:Sodium / Q000378:metabolism; D013379:Substrate Specificity
D003470:Culture Media; D007477:Ions; D009124:Muscle Proteins; D005973:Glutamine; D006639:Histidine; D001216:Asparagine; D012964:Sodium