(A) Two-state model for aptazyme function. The aptazyme conformers with low and high affinities for ligand are shown as
A and
B, respectively. The ligand-bound states of these two conformers are shown as
AL and
BL.
Kint is the equilibrium constant for the
A-to-
B transition.
Ka(A) and
Ka(B) are the association constants for the ligand (
L, shown as a blue hexgon) with the
A conformer and the
B conformer, respectively. The first-order cleavage rate constants for conformer A and conformer B are defined as
kCle(A) and
kCle(B), respectively. Under certain conditions (see text), the
AL conformer can be ignored and the model can be reduced to the enclosed green box. (B) The effect of cleavage tendency (
ω) on the performance of ligand-activated aptazymes. (D) The effect of cleavage tendency (
ω) on the performance of ligand-inhibited aptazymes. In (B) and (D) the relative ligand concentration ([
Ltot]/
Kd) is shown on the horizontal axis and the relative apparent cleavage rate constant (
kapp/
kCle) is shown on the vertical axis. The basal cleavage rate constants in the absence of ligand are shown by horizontal dashed lines. The values of

are shown as a vertical dotted lines. (C) The relationship between the cleavage tendency (
ω) of an aptazyme and the realistic ligand-dependent change in activity (

) is shown for ligand-activated aptazymes (left) and ligand-inhibited aptazymes (right). This relationship is shown for different maximum available ligand concentrations (

).