Functional characterization of the chicken cationic amino acid transporter-2 isoforms.
Utilization of lysine and arginine by mammalian skeletal muscle is due in large part to the system y(+) cationic amino acid transporters (CAT), particularly CAT-2A. The chicken CAT-2 (cCAT-2) gene is alternatively spliced to produce three isoforms (cCAT-2A/B/C). Chicken (Gallus gallus) CAT-2 isoforms were transiently and stably expressed in mammalian cell lines to determine cCAT-2 isoform cellular localization and transport properties. The cCAT-2A protein localized to the plasma membrane and the cCAT-2B protein localized to the cytoplasm juxtaposed to the plasma membrane. The cCAT-2C protein localized non-specifically throughout the cytoplasm. The cCAT-2A protein exhibited saturable transport. The K(t) of cCAT-2A for lysine using transient transfection was 2.6mM and the V(max) was 11.9pmol/mg protein/min. The K(t) of cCAT-2A for lysine using stable transfection was 7.9mM and the V(max) was 12.8pmol/mg protein/min. Transient and stable transfections of cCAT-2B or cCAT-2C resulted in no lysine or arginine transport. These data indicate that cCAT-2A is a low affinity, high velocity transporter for lysine and arginine and is the cCAT-2 isoform responsible for lysine and arginine transport in avian skeletal muscle.
Kirsch
Sandra
S
Department of Animal and Avian Sciences, University of Maryland, College Park, 20742, USA.
Humphrey
Brooke D
BD
D000818:Animals; D001120:Arginine / Q000378:metabolism; D030161:Avian Proteins / Q000032:analysis / Q000378:metabolism*; D001692:Biological Transport; D027202:Cationic Amino Acid Transporter 2 / Q000032:analysis / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D008239:Lysine / Q000378:metabolism; D020033:Protein Isoforms / Q000032:analysis / Q000378:metabolism
D030161:Avian Proteins; D027202:Cationic Amino Acid Transporter 2; D020033:Protein Isoforms; D001120:Arginine; D008239:Lysine