20.

320 Exam 2 Review Problem Solutions April 20, 2006

1.
No inhibition
vmax [ S ]
v=
K M + [S ]
vmax [ S ]
0.5vmax =
K M + [S ]
0.5 K M + 0.5[ S ] = [ S ]
[S ] = K M

Competitive
vmax [ S ]
v=
⎛ [I ] ⎞
K M ⎜1 + ⎟ + [S ]
⎝ KI ⎠
vmax [ S ]
0.5vmax =
⎛ [I ] ⎞
K M ⎜1 + ⎟ + [S ]
⎝ KI ⎠
⎛ [I ] ⎞
0.5K M ⎜1 + ⎟ + 0.5[ S ] = [ S ]
⎝ KI ⎠
⎛ [I ] ⎞
K M ⎜1 + ⎟ = [S ]
⎝ KI ⎠
⎛ [I ] ⎞
∴ K M ,CI = K M ⎜1 + ⎟
⎝ KI ⎠

Non-Competitive
⎛ ⎞
⎜ v [S ] ⎟
⎜ max ⎟
⎜ 1 + [I ] ⎟
⎜ K I ⎟⎠
v=⎝
K M + [S ]
⎛ ⎞
⎜ v [S ] ⎟
⎜ max ⎟
⎜ 1 + [I ] ⎟
⎜ K I ⎟⎠
0.5 max = ⎝
v
[ I ] KM + [S ]
1+
KI
0.5 K M + 0.5[ S ] = [ S ]
0.5 K M = 0.5[ S ]
K M , NI = K M
20.320 Exam 2 Review Problem Solutions April 20, 2006

2.
a.
k2 k3 [ S ][ E ]0

v=
( −2 + k2 + k3 )
k
k−1k−2 + k−1k3 + k2 k3
+ [S ]
k1 ( k−2 + k2 + k3 )
k−1k−2 + k−1k3 + k2 k3
KM =
k1 ( k−2 + k2 + k3 )
k 2 k3
kcat =
( k−2 + k2 + k3 )

b.
1) Assume k3 >> k2
2) k-2 Æ 0 (irreversible)

3. EC50 would likely be greater for a competitive inhibitor because they have to compete
with the substrate whereas a non-competitive inhibitor does not.

4.
a. True
b. False

5.
ρ [ R ]0
[ L]0 
N AV

6.
a.
1) degradation
2) recycling

b.
1) degradation
2) trancytosis

7.
a.
K D , AB = K D , A ⋅ KU , B = 10−4 ⋅10−3 = 10−7

b.
K D , AB 10−7
KU , A = = −5 = 10−2
K D,B 10

c.
K D, A 10−4
[ A]eff = = −2 = 10−2
KU , A 10
20.320 Exam 2 Review Problem Solutions April 20, 2006

d.
1
KU , A

e. KD,AB increases (lower binding affinity) if linker is shorter than needed, and also
if linker is long and very flexible. KD,AB decreases (higher binding affinity) if
linker is rigid and "just the right length" to reach the 2nd binding site.

8. d
a.

160
140
120
100
no inhibitor
80 10^-5 inhibitor
60
40
20
0
0 0.0005 0.001 0.0015 0.002 0.0025

The inhibitor is a non-competitive inhibitor.

Use Lineweaver-Burke plot to analyze Vmax, KI, and KM.

b.
Use non-competitive inhibitor equation.

9.
dRs
= − k f Rs L + kr Cs − ker Rs + f xr k xr Ri + S R
dt
dCs
= k f Rs L − kr Cs − kec Cs + f xc k xc Ci
dt
dRi
= ker Rs − f xr k xr Ri − (1 − f xr ) k xr Ri
dt
dCi
= kecCs − f xc k xcCi − (1 − f xc ) k xc Ci
dt
R0 = Ri + Rs + Ci + Cs
Cs ρ Ci ρ
L0 = L + +
N AV N AV