Consider a
molecule that interconverts between two chemical states B and A:
Then the chemical
equilibrium expression Keq can be described as:
If a
molecule S binds to both B and A forming SB and SA,
then we have the following system:
The initial
binding of S will perturb the initial equilibrium concentrations
of B and A by the affinity constants K1 and K2 that S has
for A and B respectively. Under these initial
conditions, the amounts of SB and SA are
given by the Langmuir binding expressions:
and
If then
the binding of S to A and B will
depend on the initial concentrations of [A] and [B] and won't
perturb the initial ratio of [A]/[B]. If ,
then the initial binding of S to A and B will
be relatively unequal, which will perturb [A]/[B]. This
relatively unequal binding will change the initial equilibrium as originally
expressed by Le Chatelier in his famous principle. The stress on the original
equilibrium from the binding of S will deplete one side of the
equilibrium (given ),
which would then be compensated by a shift toward the depleted species to
restore the equilibrium. This shift would necessitate the transfer of some
amount of the species from the other side of the equilibrium to the relatively
depleted side of the equilibrium.
As an
aside, if we weren't aware that S was binding then we would
write the equilibrium expression simply as [A]/[B]. However,
if S was present and binding preferentially to B,
for example, then the equilibrium concentration of the B species
would increase by Le Chatelier's principle because the total B species
would include the free B plus SB plus the
amount shifted from species A to relieve the stress on the
original equilibrium so that
the total amount of the B species would increase relative to
what it was initially. Also, the equilibrium concentration of species A would
be equivalently decreased by the conservation of matter law. The net effect
would appear to shrink the equilibrium constant Keq in the
presence of S. However, in fact the equilibrium constant doesn't
really change for the free species concentrations of A and B.
It only appears to change because we are now including the shift from A into
our chemical notation for the concentration of species B. The
origin of this shift is the relatively unequal binding of S to A and B.
Although there has been much previous thought about how to calculate this shift,
the present analysis presents the most simple and direct method.
In order to
calculate the net perturbation or shift ()
due to the relatively unequal binding of S to B and A,
we can use the mathematically derived fundamental equation for equilibrium (link) with the following
substitutions: b = [B], f(x) = SA, a =
[A] and g(x) = SB,
and further
substituting the Langmuir binding expressions for SA and SB gives,
and further
simplifying,
we finally
get,
(3)
This
expression compares the two Langmuir binding functions for SA and SB for
their relative effects on [B] and [A] by determining within
the domain S. This allows us to understand how the binding of S simultaneously
to A and B perturbs the original chemical
equilibrium between them.
Apart from the
fact that the fundamental equation for equilibrium was mathematically derived,
an objection may be made to the fact that SA and SB were
added to A and B rather than
subtracted. However, upon reflection we see that the addition makes
more sense. If SA was subtracted from A instead
of added as in the following,
(4)
then the ratio
would approach zero as more of S binds to A. This
would reflect a decrease in the complementary numerator A + ,
which wouldn't make sense from the perspective of Le Chatelier's principle
because the increase in relative binding of S to A should
produce a greater shift in the equilibrium toward the A side
of the equilibrium and a positive shift for not
a negative shift (note that this is true for this particular example, but isn't
meant to imply that can
never be negative). Certainly this makes sense for the
straightforward case when S binds only to A and
not to B.
Therefore our
thinking must be correct in order to understand this shift within the context
of Le Chatelier's principle as applied to coupled equilibria. It may
be more correct to consider that the binding of S to A increases
the potential chemical species of A by including the formation
of a new potential species SA. Therefore this increase in the
potential chemical species should be accounted for by the addition of SA and SB to A and B rather
than their subtraction (This is similar to the ratios of probabilities in a
partition function with the disjoint probabilities being added). In this sense,
the ratio on the left side of equation (4) represents the potential chemical
species of A compared to B in the presence
of S. Any increase in the formation of SA is
considered as an increase in the potential reservoir for the A species
thereby increasing the probability that we would find more A species
given the condition that .
This approach
explains why there is a close correlation between the thermodynamic coupling
free energy,,
for an acid-base, two-state model and the experimentally determined efficacies
for ligands binding to the 5-HT2A receptor[1].
Similarly, the ratio
determined by fitting an acid-base, two-state model to the pH-dependent binding
significantly correlated with the experimental efficacies for a variety of
ligands (link). This is due to the ligand's ability to relatively favor the
base form of the receptor and thereby produce a shift in the original
equilibrium to create more of the base state of the receptor. This presents a
general mechanism for receptor activation within the confines of a plausible
biophysical model.
THE TWO-STATE
MODEL: This is similar to most other two-state models with R and R* states
corresponding to inactive and active receptor states except that this model
relates the response to a fundamental equation for physical equilibrium, which
can be solved for the net shift in the original equilibrium, RH,
Where RH and
RL represent the amount of unperturbed receptor existing in
initial high and low affinity states respectively, and D represents the
concentration of the binding drug or ligand.
With the
dissociation constants, KDH and KDL, for the high
and low affinity binding, this equation has been shown to accurately model the
dose-response behaviors for a wide variety of drug-receptor systems (see EXPERIMENTAL
VERSUS CALCULATED DOSE-RESPONSE CURVES below).
EXPERIMENTAL VERSUS CALCULATED DOSE-RESPONSE CURVES
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