Protein Interactions:

A Basis for Protein Function

Prerecorded Assignment for Class 3

Rod Hori
University of Tennessee
Introduction, Langmuir Isotherm and
Scatchard Plot
 Protein Binding Interactions are
Important in Protein Function

Examples
Myoglobin and Hemoglobin
Antibodies
Glucocorticoid Receptor
Enzymes
(Additional ones listed in supplement.)
Protein Binding Interactions are
Important in Pharmaceuticals

Examples
1. LisPro
2. Allopurinol
Binding Equilibria

We will be considering reversible

interactions
Representation of Binding

P+A PA

When it reaches equilibrium,

[P][A]
Kd =
[PA]

Kd is in the units of concentration

Lower Kd means stronger association (affinity)

Why is Affinity Important?

The efficacy of a drug is determined, in

part, on its affinity for its target.
Drug Development:
Specificity versus Affinity
 How Much Ligand Does the Protein Bind at a
Given Ligand Concentration?

Example: Ligand that inhibits enzyme

For protein with one ligand binding site:

[Pt] = [P] + [PA] Pt defined as total protein

r = average number of ligand molecules bound per

protein molecule

[PA]
r= =
[Pt]
How Much Ligand Does the Protein Bind at a
Given Ligand Concentration?
[PA]
r= (from previous slide)
[Pt]

[PA] [PA]
r= =
[Pt] [PA] + [P]
[P] [A]
Substitute into Kd =
[PA]

[A]
r= Langmuir Isotherm****
Kd + [A]
 Saturation Binding Curve

[A]
 Saturation Binding Curve

[A]
r =
Kd + [A]

r 0.5 [A]
0.5 =
Kd + [A]

=> Kd = [A]

Kd
[A]
Scatchard Plot

r 1 r

= -
[A] Kd Kd
Graphical representation of
Scatchard plot
-1
Slope =
Kd
r
[A]

r
End of this subsection