Intestinal glycyl-L-phenylalanine and L-phenylalanine transport in a euryhaline teleost.
The transport mechanisms for the dipeptide glycyl-L-phenylalanine (Gly-Phe) and L-phenylalanine (Phe) were characterized in fish intestinal brush-border membrane vesicles (BBMV). Gly-Phe was rapidly hydrolyzed only intravesicularly with almost total hydrolysis occurring even at 10 s. Dipeptide uptake was not stimulated by an inward gradient of Na, K, or H. Phe uptake was stimulated by an inward gradient of either Na or K but displayed an overshoot phenomenon only in the presence of an Na gradient. Kinetic analysis of the effect of substrate concentration on transport rate revealed that transport of both Gly-Phe and Phe occurred by a saturable process conforming to Michaelis-Menten kinetics. The Km for Gly-Phe was 9.8 +/- 3.5 mM, whereas that for Phe in the presence of Na or K, respectively, was 0.74 +/- 0.13 and 1.1 +/- 0.37 mM. Maximum uptake for Gly-Phe and for Phe in the presence of Na and K was 5.1, 0.9, and 0.4 nmol.mg and protein-1.5 s-1, respectively. Gly-Phe and Phe transport displayed different patterns of inhibition by dipeptides and amino acids. These results suggest that Gly-Phe and Phe are transported via different mechanisms, with Gly-Phe being hydrolyzed during a carrier-mediated, cation-independent process and Phe being transferred via a Na+ cotransport process similar to that described in mammals. During conditions of high luminal dipeptide concentrations, the Gly-Phe pathway may make a significant contribution to total Phe uptake.
Reshkin
S J
SJ
Department of Zoology, University of Hawaii, Manoa, Honolulu 96822.
Ahearn
G A
GA
D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D002412:Cations / Q000494:pharmacology; D004151:Dipeptides / Q000378:metabolism / Q000493:pharmacokinetics*; D005399:Fishes / Q000378:metabolism*; D006868:Hydrolysis; D007413:Intestinal Mucosa / Q000378:metabolism*; D007700:Kinetics; D008871:Microvilli / Q000378:metabolism; D010649:Phenylalanine / Q000493:pharmacokinetics*
D000596:Amino Acids; D002412:Cations; D004151:Dipeptides; D010649:Phenylalanine; C029660:glycylphenylalanine