LECTURES 30‐36: Receptor‐Ligand Binding

Problem 1

Obtain the half-time for the following ligand given the binding to the receptor and
the following conditions.

k-1=0.04 min-1 kD = 25 µM

CL0 = 2,, 20,, 100 µ

µM
Solution: The Half-time period for the receptor –ligand binding is given by
ln 2
t 1
2
=
⎛ C ⎞
k−1 ⎜1 + L0 ⎟ (8.1)
⎝ kD ⎠
For CL0 = 2 µM,
t 1
2
= 16.04 min
F CL0 = 20 µM,
For M t 1
2
= 9.627
9 627 min
i

For CL0 = 200 µ

µM, t 1
2
= 1.9254 min
Problem 2

Use the following data to determine the specific binding of a ligand A

to a receptor
p B as well as the values of kd and the number of receptors
p
per cell.

CL0, M
M Amount bound without unlabeled ligand
Amount bound without unlabeled ligand  Amount bound with 100 excess 
Amount bound with 100 excess
unlabeled ligand
1.5×10‐10 16, 000 6, 000

6× 10‐10 60 000
60, 000 24 000
24, 000

2.1×10‐9 120, 000 60, 000

7 5 10‐99
7.5×10 350 000
350, 000 260 000
260, 000

1.5×10‐8 600, 000 510, 000

Solution: The specific binding of ligand to receptor is given below.

CL0, µM Specific binding of a ligand A to receptor B (Nc)

1.5×10‐4 10, 000

6× 10‐4 36, 000

2.1×10‐3 60, 000

7.5×10‐33 90, 000

1.5×10‐2 90, 000

Calculating Nc/CL0 and after plotting Nc/CL0 Vs. Nc we get

kd= 6.67×10-10 M

NRT/kd= 7.2×107 receptors/cell/µM

Therefore NRT = 4.8×104

Problem 3: The model equation for ligand depletion is given by
du
= (1 − u )(1 − η u ) α − u (1)
dτ
NC 0
with u =u0 = at τ =0.
N RT
Solve the model equation to establish a relationship between u and τ.
τ

Solution: From equation (1), we can write

du
= dτ
⎡⎣(1 − u )(1 − η u ) α − u ⎤⎦ (11.1)

After algebraic manipulations, we get

du
= dτ
⎡⎣ηα u − (ηα + α + 1) u + α ⎤⎦
2
(11.2)
Taking out ηα common from the denominator, we get

du
= dτ (11.3)
⎡ 2
ηα ⎢u −
(ηα + α + 1) u+
1 ⎤
⎥
⎣ ηα ηα ⎦

Adjusting the terms in denominator to form quadratic equation in

terms of u,
du ((11.4))
= dτ
⎡⎛ (ηα + α + 1) ⎞ ⎛ 1 (ηα + α + 1) ⎞ ⎤
2 2

ηα ⎢⎜ u − ⎟ + ⎜⎜ − ⎟⎥
⎢⎣⎝ 2ηα ⎠ ⎝η 4η α
2 2
⎟⎥
⎠⎦
Integrating equation (11.4) on both sides
1 du
ηα ∫ ⎡⎛
= ∫ dτ (11.5)
(ηα + α + 1) ⎞ ⎛ 1 (ηα + α + 1) ⎞ ⎤
2

⎢⎜ u − ⎟ +⎜ − ⎟⎥
⎢⎣⎝ 2ηα ⎠ ⎝η 4η α
2 2
⎠ ⎥⎦

Let
⎛ 1 (ηα + α + 1) ⎞
⎜ − ⎟ = ± K 2

⎝η 4η α
2 2
⎠
Equation (11.5) becomes,
1 du
∫
ηα ⎡⎛ (ηα + α + 1) ⎞ 2
⎤
= ∫ dτ
(11.6)
⎢⎜ u − ⎟ ±K ⎥
2

⎢⎣⎝ 2ηα ⎠ ⎥⎦
 (11 6) is of the form ∫
Equation (11.6)
(
dx
)
x2 ± a2
having solution
1
a
tan( )
x +C
a

Therefore the solution of equation (11.6) is

1 ⎡⎣( 2η u + η + 1) + 1⎤⎦
τ= tan +C (11.7)
ηα K 2 Kη
Put initial condition in equation (11.7), then we obtain

1 ⎛ ⎡⎣( 2η u + η + 1) + 1⎤⎦ ⎡⎣( 2η u0 + η + 1) + 1⎤⎦ ⎞

τ= ⎜ tan − tan ⎟
⎜
ηα K ⎝ 2 Kη 2 Kη ⎟
⎠
(11.8)