Signal Transduction 1

Important Concepts in Signal Transduction

• Primary messengers
• Membrane receptors
• Second messengers
• Amplification
• Signal termination
• 7TM receptors
• G proteins
• Adenylate cyclase - Protein Kinase A
• Phospholipase C- Protein Kinase C, Ca2+
Channels
Cells respond to the
environment:

Signal Transduction
The term signal transduction
refers to the biochemical
mechanism responsible for
“transmitting” extracellular
signals inside the cell, which
ultimately lead to the activation
of target proteins that control
metabolic pathways or regulate
gene expression.

Courtesy: Roger Miesfeld

 Steps of Signal Transduction

1. Signal molecule (primary messenger, first messenger,

ligand) travels to the cell.

2. Primary messenger binds to the extracellular domain of

a receptor protein and initiates a structural change in the
receptor which is propagated across the membrane.

– Membrane receptors sense the stimulus and transfer info across

the membrane.
– Exception: some molecules, for example steroid hormones,
move across membranes, bind to proteins and act, generally at
the nucleus.
– Most molecules are too polar or too large to cross the
membrane, so the stimulus does not enter without membrane
receptors.
– Generally, receptors are intrinsic (integral) membrane proteins
with extra- and intracellular domains.
 Steps of Signal Transduction

3. Receptor protein stimulates signaling proteins

4. Second messengers amplify the signal

– Free to diffuse
– Cross talk between pathways exists

5. Second messengers bind to additional signaling proteins

6. Signal is propagated, often by a protein kinase cascade

 Steps of Signal Transduction

7. Target proteins are affected (activated, inhibited)

– Transcription factors
– Metabolic enzymes
– Cytoskeletal proteins
– Transport proteins
– Etc.

8. Signal is terminated

– Phosphatases
The biochemical basis for signal transduction
involves three primary mechanisms:

1) protein conformational changes

2) covalent protein modifications

3) altered rates of gene expression

First/primary messengers are small
diffusible biomolecules.
These can be produced through
endocrine mechanisms and act at a
distance.
Or they can function locally as
paracrine or autocrine signals.

Courtesy: Roger Miesfeld

 Small molecules act as diffusible signals

First/Primary Messengers

Human growth hormone and insulin

are peptide hormones
Cortisol is a steroid that is derived
from cholesterol
Epinephrine, also known as
adrenaline, is a derived from the
amino acid tyrosine
Acetylcholine is a neurotransmitter
that binds the acetylcholine receptor
NO is produced by deamination of L-
arginine
 Small molecules act as diffusible signals

Second Messengers

Second messengers amplify the

receptor-generated signal

Fine tuning

Rapid production of maximum response

 One of the best characterized second messengers
is cyclic AMP (cAMP).
Produced by the enzyme
adenylate cyclase from ATP.
Receptor activation of
adenylate (adenylyl) cyclase
generates large amounts of
cAMP, which in turn, binds to
and activates downstream
signaling proteins such as
cAMP dependent protein
kinase A (PKA). Importantly,
the intracellular concentration
of cAMP is carefully controlled
by the relative levels of
receptor-activated adenylate
cyclase and soluble forms of
cAMP phosphodiesterase
(PDE) which converts cAMP to Courtesy: Roger Miesfeld
AMP.
The intracellular concentration of cAMP is carefully controlled

Relative levels of receptor-activated adenylate cyclase and soluble forms of cAMP

phosphodiesterase (PDE) which converts cAMP to AMP.

Courtesy: Roger Miesfeld

 Another second messenger important in signal
transduction is cyclic GMP (cGMP)
Produced from GTP by the enzyme guanylyl cyclase. The cGMP analog Sildenafil,
also know as Viagra, is used to treat sexual dysfunction by inhibiting the activity of
cGMP phosphodiesterase (PDE). The molecular structure of sildenafil is similar to
cGMP and binds tightly to cGMP PDE.

Courtesy: Roger Miesfeld

The Kissing Bug, Rhodnius prolixus, delivers NO to their victims by injecting
heme-containing proteins called nitrophorins that carry NO into the wound along
with their saliva.

Courtesy: Roger Miesfeld

 Some Other Second Messengers: diacylglycerol (DAG),
inositol 1,4,5-trisphosphate (IP3) and calcium ion (Ca2+).

Intracellular levels of DAG, IP3 and Ca2+ are

controlled by the activity of a membrane
associated enzyme called phospholipase C
(PLC).

Receptor-mediated activation of
phospholipase C leads to cleavage of the
membrane phospholipid phosphatidylinositol
4,5-bisphosphate (PIP2) to form DAG and
IP3.

Courtesy: Roger Miesfeld

The term signal transduction
refers to the biochemical
mechanism responsible for
“transmitting” extracellular
signals inside the cell, which
ultimately lead to the activation
of target proteins that control
metabolic pathways or regulate
gene expression.

Courtesy: Roger Miesfeld

5 Major classes of receptor proteins:
the gatekeepers of the cell

Courtesy: Roger Miesfeld

 Seven-Transmembrane-Helix Receptors (7TM receptors)

All seven-transmembrane-helix receptors are

coupled to G Proteins :
G Protein Coupled Receptors or GPCRs

Fig. 14-4
β-Adrenergic receptor signal transduction pathway

Binding of ligand on the extracellular domain of

the β- adrenergic receptor causes a structural
change on the cytoplasmic side

This structural change causes a change in an

associated signal-coupling protein called a G-
protein (guanyl nucleotide binding protein)

The change in the G-protein involves GDP vs.

GTP binding affinity.

The G-protein activates adenylate cyclase,

increasing cAMP levels
Inactive form of G protein heterotrimer binds GDP

Notice the three different

subunits here
G Protein Activation

Receptor conformational
change causes structural
change in the G protein –
GDP leaves, GTP binds
GTP binding cause a
structural change in the G
protein—molecular switch

βγ dimer then dissociates

and activated α subunit goes
off to affect other proteins
The human genome contains 15 α, 5 β and 10 γ subunits,
leading to many different possible combinations (~1000)
and functions
The activated Gα subunit binds to other proteins to activate
them

Gαs binds to adenylate cyclase, activating it, leading to increased levels of cAMP,
which turns on Protein Kinase A.

Fig. 14-7
Examples from the adrenergic systems
Glycogen is a storage form of glucose.

Glycogen is broken down to glucose when the body needs energy.

Glucose levels in the blood are tightly controlled.

Glycogen synthesis (anabolism) and degradation (catabolism) are

highly regulated by hormone signaling.

The liver is one of two major storage depots for glycogen, which in
response to epinephrine or glucagon signaling, provides an important
source of glucose for export throughout the body when dietary
glucose is limiting.

Epinephrine is the fight or flight hormone.

Glucagon is released by the pancreas and has been called the

hunger hormone because it signals low blood glucose levels.
Examples from the adrenergic systems
Switch II, undergoes a conformational change in the presence of
GTP and is critical for stimulation of adenylate cyclase activity
 How does cAMP binding activate the
phosphorylating function of protein kinase A?
Stimulation of PKA
signaling events in
liver cells by
epinephrine
Stimulation of PKA in liver
cells by epinephrine
Stimulation of PKA
signaling events in
liver cells by
epinephrine
 Activation of phospholipase C

Human liver cells contain α1 adrenergic receptors

bind epinephrine and signal glycogen degradation through a
second messenger pathway linked to a phosphorylation cascade.

α1 adrenergic receptors are coupled to a heterotrimeric G

protein containing Gαq
activates the enzyme phospholipase C (PLC) through a
mechanism very similar to Gαs stimulation of adenylate cyclase
activity

Phospholipase C is a membrane associated protein

catalyzes the hydrolysis of phosphatidylinositol 4,5-bisphosphate
(PIP2) to form the second messengers DAG and IP3.
PLC produces second messengers DAG and IP3
Function of second messengers IP3 & DAG
IP3 binds to calcium channels on the endoplasmic reticulum
• causes a rapid increase in intracellular Ca2+ levels.

Released Ca2+ binds to protein kinase C (PKC)

• stimulates its association with DAG at the plasma membrane
resulting in activation of the PKC kinase function

Ca2+ binds to calmodulin, activating:

phosphorylase kinase
calmodulin dependent kinase

PKC and calmodulin dependent kinase

phosphorylate and inactivate glycogen synthase

Calmodulin-activated phosphorylase kinase stimulates

glycogen degradation by activating glycogen phosphorylase
Stimulation of PKC signaling events in liver cells by
epinephrine
 G proteins reset themselves

Possesses GTPase activity The βγ dimer then re-associates

with the α subunit, preventing it
from further propagating the signal

Ready to start all over again.

Fig. 14-9
Don’t forget the 7TM receptor,
it must be reset as well,
or it activates more G protein

Fig. 14-10
 Summary of G protein coupled receptor signaling

1. Receptor-mediated activation of GDP-GTP exchange in Gα subunits.

2. Gα stimulation of an effector enzyme that generates 2nd messengers.

3. Activation of a phosphorylation cascade by 2nd messenger signaling.

4. Inactivation of Gα by effector stimulation of the intrinsic GTPase

activity.

5. Signal duration is controlled by loss of 2nd messengers and receptor

desensitization.
 What happens when receptors aren’t reset?

Dopamine receptor and cocaine / amphetamines

Opiate receptor and heroin
Serotonin 5-HT1A receptor and MDMA (Ecstasy)

These are all G protein coupled receptors (GPCRs)!

First you get high, then you become addicted.

Why? Because the dopamine D2 receptor and the opiate and the 5-
HT1A receptors bind these drugs much more tightly than they do their
natural ligands. They don’t get reset properly, the whole signaling
cascade downstream is “messed up”.
After exposure, more receptor is needed to get the “normal”
physiological response. The body responds by altering gene
expression and receptor levels in the brain.

One exposure to MDMA or Meth permanently alters brain function!

There is no going back. This is a huge social problem right now.