Unified modeling of the mammalian and fish proton-dependent oligopeptide transporter PepT1.
Electrophysiological and biophysical analyses were used to compare the partial and complete transport cycles of the intestinal oligopeptide transporter PepT1 among three species (seabass, zebrafish and rabbit). On the whole, the presteady-state currents of the fish transporters were similar to each other. Rabbit PepT1 differed from the fish transporters by having slower-decaying currents, and the charge vs. potential (Q/V) and time constant vs. potential (τ/V) curves shifted to more positive potentials. All of the isoforms were similarly affected by external pH, showing acidity-induced slowing of the transients and positive shifts in the Q/V and τ/V curves. Analysis of the pH-dependence of the unidirectional rates of the intramembrane charge movement suggested that external protonation of the protein limits the speed of this process in both directions. The complete cycle of the transporter was studied using the neutral dipeptide Gly-Gln. Michaelis-Menten analysis confirmed that, in all species, acidity significantly increases the apparent affinity for the substrate but does not strongly impact maximal transport current. Simulations using a kinetic model incorporating the new findings showed good agreement with experimental data for all three species, both with respect to the presteady-state and the transport currents.
Laboratory of Cellular and Molecular Physiology, Department of Biotechnology and Molecular Sciences, University of Insubria, Varese, Italy.
Maria Daniela
MD
Renna
Rachele
R
Sangaletti
Elena
E
Bossi
Francesca
F
Cherubino
Gabor
G
Kottra
Antonio
A
Peres
D000818:Animals; D001492:Bass; D001692:Biological Transport; D003198:Computer Simulation; D004151:Dipeptides / Q000378:metabolism*; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008954:Models, Biological; D000074011:Peptide Transporter 1; D011817:Rabbits; D027981:Symporters / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000378:metabolism
D004151:Dipeptides; D029941:Fish Proteins; D000074011:Peptide Transporter 1; D027981:Symporters; D029961:Zebrafish Proteins; C000613207:slc15a1b protein, zebrafish; C039788:glycylglutamine