3

Introduction to Steady-State
Systems

In the first two chapters we have discussed certain topics that are fundamental
for equilibrium systems, especially binding. In this chapter we introduce
simple steady-state systems. Chapters 4 and 5 will continue with somewhat
more complicated equilibrium and steady-state systems, but without explicit
introduction of interaction free energies. Then, in Parts II and ill, we turn to
the main theme of the book: systems of both classes comprised of subsystems
that interact with each other, with the interactions treated explicitly.

8. Dlustrative Steady-State Systems

The purpose of this section is to introduce the reader to a few simple and
explicit models of steady-state systems, and to relate these models to the
corresponding equilibrium systems as limiting cases. In each example, the
rate constants are specified and related to the thermodynamic force. After this
introduction, in Sections 9 and 10, we shall derive the fundamental properties
of any independent steady-state system that can be represented by two-state
and three-state cycles. These results, especially on two-state cycles, will be
needed repeatedly in the remainder of the book.

One-Site Membrane Transport

The first and simplest model is introduced in Fig. 3-1, and the corresponding
two-state kinetic cycle is shown in Fig. 3-2(a). A protein molecule E is
present in a membrane (mem in Fig. 3-1) that separates two solutions, A and
B, both of which contain a ligand L at concentration CA and CB, respectively.
The ligand can bind onto E to form EL, either from side A or from side B.
Thus E can exist in the two states, E and EL. The possible on and off

T. L. Hill, Cooperativity Theory in Biochemistry

© Springer-Verlag New York Inc. 1985
44 Introduction to Steady-State Systems

Side A mem Side B

L on:
L at c A

fr ---f-:~
• L at cB

~+--i3'
(a)

(b)

L off:
Fig. 3-1. (a) Binding ofligand L on E in
fr' --+-:=~ ~:+-- i3
membrane, from either side. (b) Physical
significance of rate constants.

transitions for L, with first-order rate constants, are indicated in Fig. 3-1(b).
The cycle in Fig. 3-2(a) connects the two states of E and contains the same
rate constants.
The usual dominant direction of cyclic activity in Fig. 3-2(a) is counter-
clockwise. This is arbitrarily called the positive direction (for force and flux,
defined below). This corresponds to net transport of L in the direction
A ~ B. The rate constants in this direction are a and {3; the inverse transi-
tions have primed rate constants, a' and f3'. The binding, or on, rate con-
stants are a and {3'; the off rate constants are a' and {3. All of these are
first-order rate constants. That is, a and f3' (binding) are pseudo first order:
a = a*cA and {3' = {3*CB, where a* and (3* are second-order binding rate
constants.
When CA > CB, there will be net transport of L from side A to side B, via
binding of L on E, that is, using net cyclic activity in Fig. 3-2(a) in the
positive direction. If CA < CB, the net transport will be in the opposite direc-
tion. If CA = CB, there is no net transport: the system will be at equilibrium.
We have been referring to the activity of a single molecule E in the
membrane. Ordinarily, of course, there will be a large ensemble of indepen-
dent E molecules in the membrane. Our equations below usually refer to

Side
B

EL

H
E
Fig. 3-2. (a) Two-state kinetic cycle corre-
(b)
sponding to model in Fig. 3-1. (b) Contrast
EL with equilibrium binding of L on E.