An Introduction to Medicinal

Chemistry
Chapter 5
DRUG TARGETS
SIGNAL TRANSDUCTION
Salih Al-Jabour,Dr.rar.nat
Pharmacy Department
AlQuds University
Jerusalem, Palesinte
1. Signal Transduction involving Gs-Proteins

GS-Protein - membrane-bound protein of 3 subunits (α, β, γ)

- αs subunit has binding site for GDP
- GDP bound non-covalently

β γ
α
GDP
1. Signal Transduction involving Gs-Proteins
1.1 Interaction of receptor with Gs-protein
Ligand

Cell Ligand G-
membrane binding protein
Recepto binds
r γ ß Induced γ ß Induced γ ß
fit fit for
α α α
G-
protein
G Protein GDP GTP
G-protein alters shape
Binding site for G-protein opens
GDP binding site distorted
GDP binding weakened
GDP departs
1. Signal Transduction involving Gs-Proteins
1.1 Interaction of receptor with Gs-protein

γ ß γ ß
GTP binds Fragmentation
α α and release γ ß α

Induced fit
Binding site recognises GTP G-Protein alters
shape
Complex destablised

Notes:
Process repeated for as long as ligand bound to receptor
Signal amplification - several G-proteins activated by one ligand
αs Subunit carries message to next stage
1. Signal Transduction involving Gs-Proteins
1.2 Interaction of αs with adenylate cyclase
Notes:

Several 100 ATP molecules converted before αs-GTP deactivated

Represents another signal amplification
Cyclic AMP becomes next messenger (secondary messenger)
Cyclic AMP enters cell cytoplasm with message
1. Signal Transduction involving Gs-Proteins
1.3 Interaction of cyclic AMP with protein kinase A (PKA)
Notes:
Protein kinase A is a serine-threonine kinase
Activated by cyclic AMP
Catalyses phosphorylation of serine and threonine residues on protein substrates
Phosphate unit provided by ATP
1. Signal Transduction involving Gs-Proteins
1.3 Interaction of cyclic AMP with protein kinase A (PKA)

Adenyla
te
cyclase

AT cyclic
P AMP
Activation

Protein
Kinase A
P

Enzyme Enzym
(inactive e
) (active
Enzyme-catalysed
) reaction
1. Signal Transduction involving Gs-Proteins
1.3 Interaction of cyclic AMP with protein kinase A (PKA)

Protein kinase A - 4 protein subunits

- 2 regulatory subunits (R) and 2 catalytic subunits (C)
cAM
P C Catalytic subunit

C
R R
R R
cAMP
C
binding
C
sites Catalytic subunit
Note:• Cyclic AMP binds to PKA
• Induced fit destabilises complex
• Catalytic units released and activated
1. Signal Transduction involving Gs-Proteins
1.3 Interaction of cyclic AMP with protein kinase A (PKA)

P
Protein Protein
+ ATP + ADP

Notes:
• Phosphorylation of other proteins and enzymes
• Signal continued by phosphorylated proteins
• Further signal amplification
1. Signal Transduction involving Gs-Proteins
1.4 Glycogen Metabolism – triggered by adrenaline in liver cells
Adrenaline
β-Adrenoreceptor adenylate
cyclase
1. Signal Transduction involving Gs-Proteins
1.4 Glycogen Metabolism – triggered by adrenaline in liver cells

Notes
:
Coordinated effect: activation of glycogen metabolism
inhibition of glycogen synthesis

Adrenaline has different effects on different cells e.g.

activates fat metabolism in fat cells
 1. Signal Transduction involving Gs-Proteins
1.5 Drugs interacting with cyclic AMP signal transduction

• Theophylline and caffeine - inhibit phosphodiesterases

- phosphodiesterases responsible for metabolising cyclic AMP
- cyclic AMP activity prolonged
1. Signal Transduction involving Gi-Proteins
Notes:

• Bind to different receptors from those used by Gs proteins

• Mechanism of activation is identical

• αI subunit binds to adenylate cyclase and inhibits it

• Adenylate cyclase is under dual control (brake/accelerator)

• Background activity due to constant levels of αs and αi

• Overall effect depends on dominant alpha subunit

• Dominant alpha subunit depends on receptors activated

 3. Phosphorylation Reactions
• Prevalent in activation and deactivation of enzymes
• Phosphorylation radically alters intramolecular binding
• Results in altered conformations

NH3 NH3
NH3

O
O P O

O O
O H O P O
O
O O O

Active site
Active site ope
closed n
 4. Signal Transduction involving Gq Proteins
4.1 Interaction with phospholipase C (PLC)

Notes:

• Gq proteins - interact with different receptors from those recognised by GS and GI

• Split by the same mechanism to give an αq subunit

• The αq subunit activates or deactivates PLC (membrane bound enzyme)

• Reaction catalysed for as long as αq bound - signal amplification

• Brake and accelerator effect

4. Signal Transduction involving Gq Proteins
4.1 Interaction with phospholipase C (PLC)

Active site Active site

(closed) (open)
DG
α α α PIP 2
PLC PLC PLC

IP3

Active site
αq departs (closed)
DG
α PIP 2 α
PLC PLC

Phosphate IP 3 Enzyme
Binding deactivate
weakene d
d
4. Signal Transduction involving Gq Proteins
4.1 Interaction with phospholipase C (PLC)

Reaction catalysed

Inositol triphosphate Diacylglycerol

(polar and moves (remains in membrane)
into cell cytoplasm)
Phosphatidylinositol diphosphate
(integral part of cell membrane)

R= long chain hydrocarbons P = PO 32-

4. Signal Transduction involving Gq Proteins
4.2 Action of diacylglycerol
Notes:• Activates protein kinase C (PKC)
• PKC moves from cytoplasm to membrane
• PKC phosphorylates Ser & Thr residues of protein substrates
• PKC activates enzymes to catalyse intracellular reactions
• Linked to inflammation, tumour propagation, smooth muscle activity etc

Cell
membrane
Binding DG
DG
site for DG DG
PKC
PKC
Enzyme Enzyme

PKC Active site (inactive) (active)

closed Enzyme-
catalysed
Cytoplasm Cytoplasm reaction
Cytoplasm
DG binds to DG
PKC moves to Induced fit
binding site
membrane opens active site
4. Signal Transduction involving Gq Proteins
4.4 Action of inositol triphosphate

Notes:

• IP3 is hydrophilic and enters the cell cytoplasm

• Mobilises Ca2+ release in cells by opening Ca2+ ion channels

• Ca2+ activates protein kinases

• Protein kinases activate intracellular enzymes

• Cell chemistry is altered leading to a biological effect

4. Signal Transduction involving Gq Proteins
4.4 Action of inositol triphosphate
Cell
membrane

IP
3
Cytoplasm

Calmodulin
Calcium
Ca++ Calmodulin Ca++
stores
Activation Activation
Protein Protein
kinase P kinase
P
Enzyme Enzyme Enzyme Enzyme
(inactive) (active) (inactive) (active)

Enzyme-catalysed Enzyme-catalysed
4. Signal Transduction involving Gq Proteins
4.5 Resynthesis of PIP2

Several
steps
IP 3 + DG PIP 2

Inhibition

Li+ salts

Lithium salts used vs manic depression

5. Signal Transduction - Tyr Kinase Linked Receptors
5.1 Reaction catalysed by Tyrosine Kinase

O Tyrosine O
H H
N C kinase N C
Protein Protein Mg+ Protein Protein
+

ATP ADP
OH O P

Tyrosine Phosphorylated
residue tyrosine residue
5. Signal Transduction - Tyr Kinase Linked Receptors
5.2 Activation and phosphorylation of receptor

Ligand Ligand

P P
P P
P P
P P P P

Signalling protein
5. Signal Transduction - Tyr Kinase Linked Receptors
5.2 Activation and phosphorylation of receptor

Notes:
Active site on one half of dimer catalyses phosphorylation of tyrosine residues on other
half

Dimerisation of receptor is crucial

Phosphorylated regions act as binding sites for further proteins and enzymes

Results in activation of signalling proteins and enzymes

Message carried into cell

5. Signal Transduction - Tyr Kinase Linked Receptors
5.3 Signalling pathways 1-TM
Receptors
Tyrosine kinase
Guanylate cyclase
inherent or associated

Signalling proteins cGM

P

PLC IP 3 GAP Grb2 Others

γ kinase

IP 3 DG PIP 3

Ca ++ PKC
 5. Signal Transduction - Tyr Kinase Linked Receptors
5.4 Example of a signalling pathway
Growt
h
factor

1) Binding of
growth factor
Dimerisation Phosphorylation
2) Conformational
change

H H H H PO PO
O O O O O OO O O O
H OH H OH H OH OH OH PO PO PPO OP
O H O H O H H P

Binding
Grb2 SoS
Ras and

Grb2 SoS Ras GDP

Binding of Grb2 and GTP/GDP Grb2 SoS
SoS exchange
GTP
PO PO O O
P PO PO
O O P
PO PO PPO O PO POPPO O
P
5. Signal Transduction - Tyr Kinase Linked Receptors
5.4 Example of a signalling pathway

Ras
Grb2 SoS Gene transcription
PO O
PO
OP P
PO PO PO O
P

Raf (inactive) Raf (active)

Mek (inactive) Mek

(active)
Map kinase (inactive) Map kinase (active)

Transcription Transcription