FSH

FSHR

LH

LHR

Gs

Gi

Gq

Gh

EGFR

Src

Transactivation

PLCδ

IP3

cAMP

PKAERK EPAC PI3K

Aktp38

Cdk4

Appl1

ßarr

Rps6

PDE

Figure 1

k0

k1

Ordinary differential equation (ODE)

Mass action law:

k0

k1Mass action law:

(1)

(2)

(3)

A

B

A B

A B

C

k0

k1

k2 k3

k4

C DA B

C D E

F G

[B]

Time

Simulation

δ1

δ2

δ3

δ4δ5

Experimentalpoints

E Initial parameter values (k0, ...,kn)

Simulation

Compute errorChange (k0,..., kn)

Done !

Error=0 ?

δ0B

B

B

B

BB

error =f(δ0, δ1, δ2, δ3, δ4, δ5 )B B B B B B B

pER

K (

µm

ol/L

)

[Ang]

Figure 3

k0k1k2k6

k3

k7

k4

k8

k10

k11 k13 k12 k14 k20 k24 k19 k23

k17

k18

k5

k9

k22k21

k25

k16

k15

Model equations Conservation laws

Biological conditions• Condition 1: control• Condition 2: PKC inhibition, k29 k33• Condition 3: siRNA Barr2, k31 k34• Condition 4: siRNA GRK23, k10 k35• Condition 5: siRNA GRK56, k18 k36

A

B

C

[C]

Time

Stimulation

A B

Time

Stimulation

[C]

Figure 4

Epac

RFP GFPEpac

RFP GFP

BRET

cAMP

BRET

A

BRETBRET

Rluc

Gα

Ven

Gß/γ

Rluc

Gα

Ven

Gß/γ

B

BRET

cha

nge

Time (min)

FSH

Time (min)

LH

Time (min) Time (min)

GFP

BRET

C

AEQ

Ca2+

GFPAEQ

D

BRETßarryPet

Rluc Rlucßarr

yPet

+hFSH +hCGTime (s) Time (s)

Time (min) Time (min)

+hFSH +hCG

Figure 5

% o

f res

pons

e

log([hFSH]) log([hCG])

Figure 6

Effector

Kd

τ

Ligand

[Ligand]

[Effector]

Emax

EC50

A

Kd

Ligand

B

Effector1 Effector2

[Ligand]

[Effector]

Emax-1

EC50

-1

Emax-2

EC50

-2

Kd

Ligand

C

Effector1 I1

Effector2

τ1 τ2

Kd

FSH

D

Gαs ß-arrestin

τ(G,FSH) τ(ß,FSH)

Kd

L1

Gαs ß-arrestin

τ(G,L1) τ(ß,L1)

τ(ß,L1)τ(G,L1)

τ(ß,FSH)τ(G,FSH)>

Kd

L2

Gαs ß-arrestin

τ(G,L1) τ(ß,L1)

τ(ß,L2)τ(G,L2)

τ(ß,FSH)τ(G,FSH)<

Figure 7

Time (min)

activ

e G

(µm

ol/L

)

AT1-

ß-ar

rest

in 2

sig

nalin

g co

mpl

ex (µ

mol

/L)

Time (min)

Figure 8