CELL SIGNALING 3

Second messengers are the intracelluar signals which

serve as downstream signaling elements that
propagate and amplify the signals initiated by ligand
binding.

Second messenger molecules are chemically diverse e.g.

inorganic ion (Ca2+)
cyclic nucleotides (cAMP, cGMP)
lipids (diacylglycerol; DAG)
gas (nitric oxide)

 Different second messengers function in discrete subcellular compartments e.g. Ca2+ in cytosol or diacylglycerol
(DAG) in lipid bilayer.
Messages are encoded by the change in concentration of
the second messenger (rise or fall).
Second messengers convey signals by binding to, and
altering the conformation and behavior of, selected
signaling molecules and effector proteins.

Targets for activated trimeric Gproteins

The major targets for activated trimeric G-proteins

adenylyl cyclase
phospholipase C
K+ ion channel

Adenylyl cyclase - an effector enzyme that produces the second

messenger cAMP

Adenylyl cyclase (AC) is a large,

multiple transmembrane domain
protein.
Catalytic domain is located on
cytosolic face of membrane.
Gs stimulates AC activity, while
Gi forms of GPCR inhibits AC
Gs subunit stimulates AC by
mediating a conformational change
in catalytic domains of AC

cAMP pathway
cAMP is produced from ATP via action of adenylyl cyclase

HOW DOES cAMP THEN ACT?

It activates protein kinase A

Active protein kinase A then acts on

serine residues on target protein

Activation of protein kinase A (PKA) by cAMP

In some cells increase in cyclic AMP activates the

transcription of specific target genes that contain a
regulator sequence called cAMP response element

 Signal carried from cytoplasm to

nucleus by catalytic subunit of
protein kinase A
Protein kinase then
phosphorylates a transcription
factor called CREB
Leads to coactivators and
transcription of cAMP inducible
genes.

Activation of target gene expression via PKA / CREB

CRE - cAMP response element; CREB protein that binds to CRE sequences.

Note - PKA can have many additional effects on the cell, not
just CREB !
The targets for PKA differ depending upon the specific cell
type.

When cAMP is elevated in skeletal muscle and liver cells, the

breakdown of glycogen is stimulated.

In cardiac muscle, the elevation of cAMP strengthens heart

contraction, whereas in smooth muscle contraction is inhibited.

In intestinal epithelial cells, it causes the secretion of salts and

water into the lumen of the gut.

REGULATION OF PROTEIN
PHOSPHORYLATION

 cAMP can directly regulate ion channels independent

of protein phosphorylation.
This pathway is involved in sensing smell
When it directs the opening of the Na+ channels it
causes depolarization of membrane & initiation of
nerve impulse

cGMP
Guanylyl cyclases are activated by NO, CO and
peptide ligands.
Guanylyl cyclase increases the level of cGMP
cGMP then mediates its action through cGMP
dependent protein kinases.

Two cyclic nucleotide monophosphates

adenosine 3',5'-cyclic monophosphate (cAMP)
guanosine 3',5'-cyclic monophosphate (cGMP)

Page 19

 Well characterized role of cGMP is seen in the

vertebrate eye.
Important for converting visual signals received as
light to nerve signals.

 Phototransduction is the process through which photons,

elementary particles of light, are converted into electrical
signals.
Visual phototransduction occurs in the retina through
photoreceptors, cells that are sensitive to light.
Both rods and cones contain opsin, a G protein-coupled
receptor.
Opsin is bound to a light-absorbing chromophore, 11-cisretinal
Rhodopsin is present in rods and transduces dim light while
photopsins are present in cones and generate color vision

Regulation of cGMP phosphodiesterase - vision depends on a GPCR that

regulates a cyclic nucleotide-gated ion channel.

PHOSPHOLIPIDS &Ca2+
Phosphatidyl inositol 4,5 bisphosphate is known as
PIP2
This is a present on the inner leaflet of the plasma
membrane.
PIP2 can be hydrolyzed by phospholipase C
This producers two distinct second messengers called
DAG and IP3

What activates the hydrolysis if PIP2?

G protein coupled receptors PLC -
Protein tyrosine kinases

PLC -

 IP3 is a small polar molecule that is released into the

cytosol where it acts to signal the release of Ca2+
from intracellular stores

 Calcium level in the cell then increases from 0.1

to
This affects the activities of various target proteins
including kinases and phosphatatses.

How does Ca2+ mediate its effects?

When the concentration of Ca2+ increases to 0.5
microns, calcium binding protein CALMODULIN is
activated

Proteins activated by this complex

are CaM kinase family

Ex : myosin light chain kinase

Intersections between calcium and cAMP signaling pathway

CaM phosphorylates transcription factors --- CREB
Ca2+/ Calmodulin regulates adenlyl cyclase and
phosphodiesterase
cAMP regulates calcium channels

Thus these two pathways function coordinately to

regulate many cellular responses

Diacylglycerol
DAG activates the serine threonine kinases belonging
to protein kinase C family

Ca2+ in electrically excitable cells

Neurons triggers the release
of neurotransmitters
Muscle triggers muscle
contraction

Phospholipase C

produces two independent second

messengers, inositol 1,4,5 triphosphate
(IP3) and diacylglycerol (DAG), by
cleavage of phosphoinositol 4,5
bisphosphate (PIP2).
DAG is a hydrophobic lipid that can
translocate laterally through the inner
leaflet of the lipid bilayer.
IP3 is a hydrophilic molecule that can
diffuse rapidly through the cytosol.
IP3 can bind to IP3 receptors on surface
of ER and mediate rapid release of Ca2+.
In turn, Ca2+ and DAG each bind to, and
activate PKC.
PKC phosphorylates targets on serine
and threonine residues.

Cells contain different kinases and lipases that produce discrete signaling
molecules from phosphoinositides

Several different phospholipases (e.g. -C, -D, -A2) can cleave PIP2 at
different locations to generate different molecules that affect additional
signaling pathways (-C = DAG+IP3; -D = phosphatidic acid; -A2 =
arachidonic acid).
Arachidonic acid is important intermediate in synthesis of prostaglandins
that play important roles in inflammation.

ADRENERGIC HORMONES
When secreted into the bloodstream, epinephrine and
norepinephrine stimulate changes in many different tissues or
organs, all aimed at preparing the body for dangerous or stressful
situations
Overall, the adrenergic hormones trigger increased cardiac
output, shunting blood from the visceral organs to the muscles and
the heart, and cause dilation of arterioles to facilitate
oxygenation of the blood.
In addition, these hormones stimulate the breakdown of glycogen
to supply glucose to the muscles.
Adrenergic hormones bind to a family of G protein linked
receptors known as adrenergic receptors.

The adrenergic receptors are located on the smooth muscles

that regulate blood flow to visceral organs.
This activates Gq (trimeric G-protein), which in turn activates
phospholipase C (effector), which produces IP3 and DAG (second
messengers) from PIP2.

Contraction of smooth muscle

hence constriction of blood vessels
and reduced blood flow

 The adrenergic receptors are found on smooth

muscles associated with arterioles that feed the
heart, smooth muscles of the bronchioles in the lungs,
and skeletal muscles
It activates Gs which activates the cAMP signal
transduction pathway
Leading to relaxation of smooth muscle.

In addition, these hormones stimulate the

breakdown of glycogen to supply glucose to the
muscles