Phenylalanine transport in guinea pig jejunum. A general mechanism for organic solute and sodium cotransport.
1. Sodium-dependent phenylalanine transport by guinea pig jejunum exhibits apparently pure K-type activation kinetics where Vmaxs is constant but KT decreases as [Na+] increases. At 0, 3 and 6 mM sodium, however, the results deviate from the expected hyperbolic kinetics and give a plateau. 2. This finding is interpreted in terms of the hypothesis that the outer face of the brush border membrane contains enough Na+ to support amino acid and Na+ cotransport at essentially maximal rates, even after preincubation of the tissues in vitro for several minutes in sodium-free buffers. 3. Sodium could move dynamically into this region from tissue stores and across the paracellular pathway. Passage of NaCl directly across the brush border also seems possible by reversal of the (neutral) Na+ and Cl- cotransport system. 4. To reconcile contradictory observations obtained in different laboratories, either with intact-epithelium preparations or with isolated brush border membrane vesicles, we include a theoretical analysis of the kinetics of organic solute and Na+ cotransport. For simplicity, this analysis is limited to cases of 1/2 stoichiometry and to neutral organic solutes such as sugars and monoamino-monocarboxylic amino acids. 5. Cotransport is explained in terms of a general, allosteric mechanism involving one site for S and another for Na+. There is no preferential order for binding, but only the ternary complex S-carrier-Na+ can translocate at quantitatively significant rates (obligatory activation kinetics). Since Na+ crosses the membrane as the free cation, under physiological conditions (inside-negative membrane potential) it will move towards its position of electrical equilibrium, hence unidirectionally. This explains why, with intact-tissue preparations, solute influx exhibits Michaelis-Menten kinetics. 6. By definition, cotransport kinetics are mixed type and involve effects on both KT and Vmaxs. Macroscopic deviations from this expected behaviour can be explained in terms of quantitative differences in the values of certain dissociation constants, all within the framework of the same general mechanism. Thus, apparently pure K-type activation kinetics will be seen when both the absolute value of the constant, K'a, and the ratio between constants, Ka/Ks, are small. The reciprocal situation will be true for systems exhibiting apparently pure V-type activation kinetics.
D000818:Animals; D001692:Biological Transport; D006168:Guinea Pigs; D007583:Jejunum / Q000378:metabolism*; D007700:Kinetics; D008871:Microvilli / Q000378:metabolism; D008954:Models, Biological; D010649:Phenylalanine / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism* / Q000494:pharmacology
D010649:Phenylalanine; D012964:Sodium