Apparent charge of binding site in ion-translocating enzymes: kinetic impact.
Recently, we presented a general scope for the nonlinear electrical properties of enzymes E which catalyze translocation of a substrate S with charge number z(S) through lipid membranes (Boyd et al. J. Membr. Biol. 195:1-12, 2003). In this study, the voltage sensitivity of the enzymatic reaction cycle has been assigned to one predominant reversible reaction step, i.e. the reorientation of either E or ES in the electric field, leaving the reorientation of the alternate state (ES or E) electroneutral, respectively. With this simplification, the steady-state current-voltage relationships (IV) assumed saturation kinetics like in Michaelis-Menten systems. Here, we introduce an apparent charge number z(E) of the unoccupied binding site of the enzyme, which accounts for the impact of all charged residues in the vicinity of the physical binding site. With this more realistic concept, the occupied binding site assumes an apparent charge of z(ES) = z(E) + z(S), and IV does not saturate any more in general, but exponentially approaches infinite or zero current for large voltage displacements from equilibrium. These nonlinear characteristics are presented here explicitly. They are qualitatively explained in a mechanistic way, and are illustrated by simple examples. We also demonstrate that the correct determination of the model parameters from experimental data is still possible after incorporating z(E) and its corollaries into the previous model of enzyme-mediated ion translocation.
Gradmann
Dietrich
D
Abteilung Phytophysik der Universität, Untere Karspüle 2, 37073 Göttingen, Germany. dgradma@gwdg.de
Boyd
Carl M
CM
D001665:Binding Sites; D001692:Biological Transport; D055592:Biophysical Phenomena; D001703:Biophysics; D004798:Enzymes / Q000737:chemistry; D007473:Ion Channels; D017136:Ion Transport; D007477:Ions; D007700:Kinetics; D008956:Models, Chemical; D018408:Patch-Clamp Techniques; D013379:Substrate Specificity
D004798:Enzymes; D007473:Ion Channels; D007477:Ions