Heterogeneity of amino acid transport in horse erythrocytes: a detailed kinetic analysis of inherited transport variation.
1. Thoroughbred horses were divisible into five distinct amino acid transport subgroups on the basis of their erythrocyte permeability to L-alanine, measured uptake rates ranging from 5 to 625 mumol l cells-1 h-1 (0.2 mM-extracellular L-alanine, 37 degrees C). 2. Erythrocytes from animals belonging to the lowest L-alanine permeability subgroup (5-15 mumol l cells-1 h-1) (transport-deficient type) exhibited slow nonsaturable transport of this amino acid. In contrast, cells from horses of the four transport-positive subgroups possessed additional high-affinity (apparent L-alanine Km (Michaelis constant) congruent to 0.3 mM) and/or low-affinity (apparent L-alanine Km congruent to 13 mM) Na+-independent transport routes selective for L-neutral amino acids of intermediate size. The two transporters, designated systems asc1 and asc2, respectively, also possessed a significant affinity for dibasic amino acids. 3. Amino acid transport activity in horse erythrocytes behaved as if controlled by three co-dominant alleles (s, h and l), where s is a silent allele, and h and l code for the functional presence of systems asc1 and asc2, respectively. 4. At physiological temperature, system asc1 operated preferentially in an exchange mode. In contrast, system asc2 did not participate in exchange reactions at 37 degrees C, but did exhibit significant trans-acceleration at 25 degrees C. 5. Reduction of the incubation temperature also resulted in dramatic decreases in apparent Km and Vmax for L-alanine uptake by system asc2, whereas the effects of temperature on system asc1 were much less marked. At 5 degrees C the two transporters exhibited equivalent kinetic constants for L-alanine influx. L-Alanine uptake by transport-deficient cells was relatively insensitive to temperature. Influx by this route may represent the ground-state permeability of the lipid bilayer. 6. The effects of low temperature on system asc2 suggest a preferential impairment of the mobility of the unloaded carrier relative to that of the loaded transporter. Similarly, the different kinetic properties of systems asc1 and asc2 at physiological temperature are attributed to a difference in the mobilities of the empty carriers, this difference being minimized at 5 degrees C.
Fincham
D A
DA
Department of Biochemistry, Faculty of Medicine, Chinese University of Hong Kong, Shatin, N.T.
Mason
D K
DK
Paterson
J Y
JY
Young
J D
JD
D000409:Alanine / Q000378:metabolism; D000596:Amino Acids / Q000097:blood* / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002463:Cell Membrane Permeability; D004912:Erythrocytes / Q000378:metabolism*; D006736:Horses / Q000097:blood*; D007700:Kinetics
D000596:Amino Acids; D000409:Alanine