L-threonine transport in pig jejunal brush border membrane vesicles. Functional characterization of the unique system B in the intestinal epithelium.
Uptake and inhibitory kinetics of [3H]L-threonine were evaluated in preparations of pig jejunal brush border membrane vesicles. Uptake of [3H]L-threonine under O-trans, Na+ gradient, and O-trans, Na(+)-free conditions was best described by high affinity transport (Km < 0.01 mM) plus a nonsaturable component. The maximal velocity of transport was 3-fold greater under Na+ gradient conditions. 100 mM concentrations of all of the dipolar amino acids and 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid caused complete inhibition of [3H]L-threonine transport under Na+ gradient and Na(+)-free conditions. Imino acids, anionic amino acids, cationic amino acids, and methylamino-isobutyric acid caused significant partial inhibition of L-threonine uptake. Inhibitor concentration profiles for proline and lysine were consistent with low affinity competitive inhibition. The Ki values of alanine and phenylalanine approximated 0.2 and 0.5 mM, respectively, under both Na+ gradient and Na(+)-free conditions. These data indicate that the transport system available for L-threonine in the intestinal brush border membrane (system B) is functionally distinct from other amino acid transport systems. Comparison of kinetics parameters in the presence and absence of a Na+ gradient suggests that both partially and fully loaded forms of the carrier can function to translocate substrate and that Na+ serves to accelerate L-threonine transport by a mechanism that does not involve enhanced substrate binding.
Maenz
D D
DD
Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, Canada.
Patience
J F
JF
D000409:Alanine / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D066298:In Vitro Techniques; D007408:Intestinal Absorption / Q000187:drug effects; D007583:Jejunum / Q000378:metabolism*; D007700:Kinetics; D008239:Lysine / Q000494:pharmacology; D008564:Membrane Potentials; D008871:Microvilli / Q000378:metabolism; D010649:Phenylalanine / Q000494:pharmacology; D011392:Proline / Q000494:pharmacology; D012964:Sodium / Q000502:physiology; D013552:Swine; D013912:Threonine / Q000378:metabolism*
D013912:Threonine; D010649:Phenylalanine; D011392:Proline; D012964:Sodium; D008239:Lysine; D000409:Alanine