The kinetic mechanism of the glutamate-aspartate carrier in rat intestinal brush-border membrane vesicles: the role of potassium.
The sodium dependent transport system for L-glutamate and L-aspartate localized in the apical part of rat enterocytes has previously been kinetically characterized (Prezioso, G., and Scalera, V. (1996). Biochim. Biophys. Acta 1279, 144-148). In this paper the mechanism by which the potassium cation specifically activates the L-glutamate-sodium cotransport process is investigated. Potassium has been found to act as an activator when it is present inside the membrane vesicles, while its presence outside is ineffective, and the effect is saturable. The kinetic parameters with respect to sodium and glutamate have been compared in the presence and in the absence of the activator. The results indicate that the ordered sodium-sodium glutamate mechanism is not altered by potassium, and that the activation is probably exerted on both the rate determining steps of the transport process. It is proposed that (1) a specific binding site for potassium is present on the inside hydrophilic part of the membrane carrier, (2) the binding of the effector accelerates the intramembrane rearrangement steps of both the disodium glutamate-carrier complex and the free carrier, (3) the affinity of the carrier is lowered with respect to sodium whereas it is increased for glutamate, and (4) K+ antiport is not performed by this carrier.
Scalera
V
V
Dipartimento di Fisiologia Generale ed Ambientale, Università degli Studi di Bari, Italy. v.scalera@biologia.uniba.it
Mola
M G
MG
Prezioso
G
G
D027322:Amino Acid Transport System X-AG / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001693:Biological Transport, Active / Q000187:drug effects; D018698:Glutamic Acid / Q000378:metabolism; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008871:Microvilli / Q000187:drug effects / Q000378:metabolism*; D008954:Models, Biological; D011188:Potassium / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley
D027322:Amino Acid Transport System X-AG; D001224:Aspartic Acid; D018698:Glutamic Acid; D011188:Potassium