Presented by Silpa P Sankar

INTRODUCTION
SIGNAL TRANSDUCTION:The interaction of a receptor with its chemical messenger is only the first step in a complex chain of events involving a several secondary messenger , protein & enzymes and ultimately change the cell chemistry.  Signal transduction process results from the activation of G-protein-couple receptor & kinase receptor.  G-protein-couple receptor -30% of the marked drugs  Kinase receptor -Novel drugs(anticancer . therapy)

Receptor

A receptor is a protein molecule embedded within the cell membrane with part of its structure facing the outside of the cell.  The protein surface will be a complicated shape containing hollows, ravines, and ridges, and somewhere amidst this complicated shape.

There is an important difference between enzymes and receptors in that the chemical messenger does not undergo a chemical reaction. It fits into the binding site of the receptor protein, passes on its message and then leaves unchanged.

1. Receptor superfamilies

RESPONSE TIME

• ION CHANNEL RECEPTORS

msecs
MEMBRANE BOUND seconds

• G-PROTEIN COUPLED RECEPTORS
• KINASE LINKED RECEPTORS

minutes

• INTRACELLULAR RECEPTORS

G protein-coupled receptors (GPCRs), also known as seven-transmembrane domain receptors situated at the inner surface of the cell membrane & made up of ά, β ,γ protein sub - units
G protein-coupled receptors are found only in eukaryotic

The receptor protein is embedded within the membrane ,with the binding site for the chemical messenger on the outer surface-

The

G protein-coupled receptor is activated by an external signal in the form of a ligand or other signal mediator. This creates a conformational change in the receptor, causing activation of a G protein. Further effect depends on the type of G protein.

    


α- subunit has a guanyl nucleotides binding pocket – Guanosine diphosphate(GDP)—In resting state Types of G – protein Gs,Gi / Go , Gq /G11 Specificity Specific G- protein are recognize by specific receptor Gs & Gi - β- adrenoceptor Gq - α – subunits The G-protein's α subunit, together with the bound GTP, can then dissociate from the β and γ subunits to further affect intracellular signaling proteins or target functional proteins directly depending on the α subunit type (Gαs, Gαi/o, Gαq/11, Gα12/13).

 Specific

mechanisms  Gαs activates the cAMP-dependent pathway by stimulating the production of cAMP from ATP. This is accomplished by direct stimulation of the membrane-associated enzyme adenylate cyclase. cAMP acts as a second messenger that goes on to interact with and activate protein kinase A (PKA). PKA can then phosphorylate myriad downstream targets.

Gαi inhibits the production of cAMP from ATP.  Gαq/11 stimulates membrane-bound phospholipase C beta, which then cleaves PIP2 (a minor membrane phosphoinositol) into two second messengers, IP3 and diacylglycerol (DAG).  Gα12/13 are involved in Rho family GTPase signaling (through RhoGEF superfamily) and control cell cytoskeleton remodeling, thus regulating cell migration.  Gβγ sometimes also have active functions, e.g., coupling to L-type calcium channels.

 There

are two principal signal transduction pathways involving the G protein-coupled receptors:  the cAMP signal pathway  the Phosphatidylinositol signal pathway

 

.αs – subunit has intrinsic GTP-ase activity(catalyse the hydrolysis of its bound GTP to GDP) cAMP-activate the enzyme protein kinase A (serine-threonine kinases-which catalyse the phosphorylation of serine & threonine residues in protein substrates

NH2 N N

NH2

Adenylyl cyclase

N N O

N N

N O O O N O P O P O P O O O O O P O P HO OH

Activate c-AMP dependent protein kinases (Phosphoryl. of proteins, i.e. enzymes)

O O P O O

OH

ATP

c-ATP

Various responces (ex. metabolism, cell division)

eg; Eg:-

The function of cAMP Targeting PKA (cyclic-AMP-dependent protein kinase A)

MECHANISM


  

Binding the chemical messenger or ligand to the receptor Binding of the G-protein to the receptor – ligand complex Α-subunit binds guanosine diphosphates (GDP) resting state. Specific G-protein recognized by specific receptors. Receptor change the shape & exposed new binding site on its inner surface. GDP GTP (change the shape),release from the site (weaken the intermolecular force) α-subunit splits off from the β & γsubunits

Adrenaline

α β-adren oceptor
ATP α

cAMP Protein kinase A Inhibitor inactive

Glycogen synthase-P (INACTIVE)

Glycogen synthase-P (ACTIVE)

C

Catalytic subunit of PKA Phosphorylase Kinase-p(active)

Inhibitor-P active Phosphatase (inhibited)

Phosphorylase Kinase(inactive)

P-b Glycogen

P-a
Glucose-1 -phosphate

 Autonomic

nervous system transmission:
both the sympathetic and parasympathetic nervous systems are regulated by GPCR pathways, responsible for control of many automatic functions of the body such as blood pressure, heart rate, and digestive processes

Behavioral and mood regulation:

Gq released αq-subunit ,phospholipase C activated or deactivated. If activated phospholipase C hydrolysis of phosphatidylinositol diphosphate (PIP2)
P Eg:-Eskalithe ,Lithane ,Lithium Carbonate
IP 3 IP 2

diacylglycerol (DG) :inositol triphosphate (IP3) :-

P

P
IP Inositol

PIs

PI

PI-4

PI-4-P5
PIP

PIP 2

Lithium salt

INHIBITION
P-Phosphatase PIs-PI synthase PI-4-kinase

Ion channels are complexes made up of 5 glycoprotein subunits which traverse the cell membrane.  The central lined with polar amino acids to give a hydrophillic pore.  Lock gate should be control by a receptor protein.  When chemical messenger binds to the external site of the receptor protein ,its changes the shape.

Mechanism

 

Receptor controlling ion channels are an integral part of the ion channel. Binding of a messenger induce a change in shape, which results in the opening of the ion channel

General structure
Receptor
Binding site Messenger

Cell membrane

INDUCED FIT ‘GATING’ (ion channel opens)

Cell membrane

Five glycoprotein subunits traversing cell membrane

Cationic ion channels for K+, Na+, Ca2+ (e.g. nicotinic) = excitatory Anionic ion channels for Cl- (e.g. GABAA) = inhibitory

Transverse view of ion channel receptor
Binding sites
Ion channel
b a g a b d g a

Cell membrane

d

a

2xa, b, g, d subunits

Two ligand binding sites mainly on a-subunits

The receptor protein in the ion channel control by the glycine in the α- subunit

Structure of protein subunits (4-TM receptor subunits)
Each subunit has 4 transmembrane
Neurotransmitter binding region
H2N

Extracellular loop
CO2H

Cell membrane

TM1

TM2

TM3

TM4

Intracellular loop

Variable loop

4 Transmembrane (TM) regions (hydrophobic)

Gating
When the receptor binds a ligand , it changes shape & this has a knock on effect on the protein complex which causes the ion channel to open ,this process is called gating

Neurotransmitter binds

Induced fit at binding site

‘Domino effect’

Rotation of 2TM regions of each protein subunit
Ion flow

TM2 Cell membrane

TM2

TM2 TM2

TM2 TM2 TM2 TM2 TM2

TM2
TM2

Transverse view of TM2 subunits

TM2

Transverse view of TM2 subunits

Closed Open

Channel Gating Mechanisms
AChR: Proposed gating mechanism

Closed

Open

• Fast response measured in msec

• Ideal for transmission between nerves
• Binding of messenger leads directly to ion flows across cell membrane

• Ion flow = secondary effect (signal transduction)
• Ion concentration within cell alters • Leads to variation in cell chemistry

Typical Ion Channels with Known Structure:

K+ channel (KCSA)

Acetylcholine receptor M2 transmembrane segment

Types of ion channels:
    Simple pores (GA, GAP junctions) Substrate gated channels (Nicotinic receptor) Voltage-gated channels (K-channels) Pumps (ATP-synthase, K+,Na+-ATPase)

Voltage-gated channels

Channel which are sensitive instead to the potentional difference that exists across a cell membrane potential
These ion channels are present in the axons of excitable cells. Transmission of a signal along individual neurons & are important drug targets for local anaesthetics.

Eg:HO OH HO HC CH 2NHCH
3

Adrenalin Adrenaline

The local anaesthetics

Gramicidin A
Antibiotic peptide
Forms a pore in the cell wall of a bacteria and lets out monovalent cations (K+, Na+). [Membrane potential disappears and bacteria dies!]

15 amino acids, helical Channel is formed by a headto-head dimer

Targeting molecules for Calcium Calcium binding protein Calmodulin

G-protein linked receptors coupled to ion channels
              

Acetylcholine (muscarinic)  Adenosine & adenine nucleotides  Adrenaline & noradrenaline  Angiotensin  Bombesin  Bradykinin  Calcitonin  Cannabinoid  Chemokine  Cholecystokinin & gastrin  Dopamine  Endothelin  Galinin  GABA (GABAB)  Glutamate (quisqualate) 

Histamine 5-Hydroxytryptamine (1,2) Leukotriene Melatonin Neuropeptide Y Neurotensin Odorant peptides Opioid peptides Platelet-activating factor Prostanoid Protease-activated Tachykinins Taste receptors VIP Vasopressin and oxytocin

Ionic bond 5-10 kcal/mol, Reversible bonding
Acetylcholin
O O H N

Hydrogen bond 2-5 kcal/mol, Reversible bonding

Hydrophobic interaction 0.5-1 kcal/mol, Reversible bonding

O O H N

O

O

Anion cavity
N H

Lipophilic area H-bind acceptor
HO

O

OH

Types of receptors
Superfamily
1

Endogenous ligands
Fast neurotransmittors ex. Acetylcholine

General structures

Ligand gated ion chanels

2

Slow neurotransm. ex. noradrenalin G-Protein coupled receptors Hormones Insuline Growth factors Enzyme coupled receptors Catalytic receptors

3

4

Steroid hormones Thyreoid hormones Vitamin A, D

Cytoplasmic receptors

Conclusion

Reference
 An

introduction to medicinal chemistry –Graham L.Patrick,fourth
edition,page Burger’s medicinal chemistry& drug

discovery ,volume 1,page  Essentials of medicinal pharmacologyby K D Tripathi (5th edition)  www.wikipedia.org,www.pubmed.com