71
which can be converted to a straight line equation having the form
y mx + b by taking a double reciprocal to give:
V
e
V
EFH
- 1
kt^h j + Kr^H
kE=7T K£=T7
[I]
This equation will give a straight line when the left side of the equa
tion is plotted with respect to the reciprocal of the concentration of
soluble ligand, I. This is shown for theoretical data in Figure 21
where [i] and Kj would have the same units. From the above linear equation
a line would have a slope of Kj/( (K]-^-/k£^j)-1) and an ordinate intercept
of 1/ ((Kjyqq/K-j^O-l). The quotient from the slope divided by the inter
cept would give the value for K-j-, the dissociation constant for the E*I
complex.
Equation 16 from above can be expressed in terms of observed elu
tion volumes by substituting in the definition, V = + V0.
This substitution yields equation 19,
- v: = V
- 1 [1]
[i] + Kt
This is similar to equation 16 in which the volume term is slightly re
arranged to give a value which can be expressed directly from experimen
tally obtained values; Ve is the elution volume at for a finite concen
tration of I and VQ is the elution volume in the absence of I.
Equation 16 can be linearized by taking a double reciprocal to give,
1 = K1 + 1
rT] kE=H
VL-M KL=M VL-M kt.7th
V V
ve vo
1