Second messenger

Diacylglycerol

Presented by :-SALONI MEHTA

Department of Pharmacology
MPL– 104T
L. M. College of Pharmacy
• One of the most widespread pathways of intracellular signaling is
based on the use of second messengers derived from the membrane
phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2).

• PIP2 is a minor component of the plasma membrane, localized to the

inner leaflet of the phospholipid bilayer. 

• A number of these second messengers are derived from

phosphatidylinositol (PI).

• The inositol group in this phospholipid, which extends into the

cytosol adjacent to the membrane, can be reversibly phosphorylated
at several positions by the combined actions of various kinases and
phosphatases. These reactions yield several different membrane-
bound  phosphoinositides.
 
 PHOSPHATIDYLINOSITOL - DERIVED SECOND
MESSENGERS :
 Phosphatidylinositol (PI) is a negatively charged phospholipid and a
minor component in eukaryotic cell membranes.
 The inositol can be phosphorylated to form
o PI
o PIP PHOSPHOINOSITIDES
o PIP2
 Intracellular enzyme phospholipase C (PLC) hydrolyses PIP2 Which Is
Found In The Inner Layer of the plasma membrane. Hydrolysis of PIP2
yields two products :
o Inositol-1,4,5-triphosphate (IP3)
o Diacylglycerol (DAG)
FORMATION OF IP3 – DAG :
 IP3 PATHWAY:

• Whereas diacylglycerol remains associated with the plasma membrane,

the other second messenger produced by PIP2 cleavage that is IP3, it is
a small, polar, soluble molecule that is released into the cytosol and
bind to receptor on the endoplasmic reticulum causing the release of
Ca2+ from intracellular stores into the cytosol.
• The cytosolic concentration of Ca2+ is maintained at an extremely low
level (about 0.1 μM) as a result of Ca2+ pumps that actively export
Ca2+ from the cell.
• As a result, cytosolic Ca2+ levels increase, which affects the activities of
a variety of target proteins, including protein kinases and phosphatases
and trigger the response.
• So, some members of the protein kinase C family require Ca2+ as well
as diacylglycerol for their activation, so these protein kinases are
regulated jointly by both arms of the PIP2 signaling pathway.
1. Example :The calcium rise is needed for NF-AT (The “nuclear
factor of activated T cells”) to turn on the appropriate genes in
the nucleus.
 DAG PATHWAY:
 
• The diacylglycerol produced by hydrolysis of PIP2 activates in
presence of phospholipase C (PLC).
• DAG stimulates protein kinase C (PKC) activity by greatly increasing
the affinity of the enzyme for calcium ions.
• PKC phosphorylates specific serine and threonine residues in the
target proteins. ( known target proteins include calmodulin, the
glucose transporter, HMG – Co A reductase, CYP450 etc. )
• A good illustration of this role of protein kinase C is provided by the
action of phorbol esters, which have been studied extensively
because they promote the growth of tumors in animals.
• This tumor-promoting activity of the phorbol esters is based on their
ability to stimulate protein kinase C by acting as analogs of
diacylglycerol.
• Protein kinase C then activates other intracellular targets,
including a cascade of protein kinases known as the MAP kinase
pathway, leading to transcription factor phosphorylation, changes
in gene expression, and stimulation of cell proliferation. 
• In addition to activating PKC, diacylglycerol has a number of other
functions in the cell:
o   a source for prostaglandins
o   a precursor of the endocannabinoid -2 arachidonoylglycerol
o   an activator of a subfamily of TRPC (Transient Receptor
Potential Canonical) cation channels,
 OTHER THAN DIACYLGLYCEROL AND
IP3 MESSENGERS:
• PIP2 not only serves as the source of diacylglycerol and IP3, but is
also the starting point of a distinct second messenger pathway that
plays a key role in regulating cell survival.
• In this pathway, PIP2 is phosphorylated on the 3 position of inositol
by the enzyme phosphatidylinositide (PI) 3-kinase. Like
phospholipase C, one form of PI 3-kinase is activated by G proteins,
while a second has SH2 domains and is activated by association with
receptor protein-tyrosine kinases.
• Phosphorylation of PIP2 yields phosphatidylinositol 3,4,5-
trisphosphate (PIP3), which functions as a distinct second messenger.
A key target of PIP3, which is critical for signaling cell survival, is a
protein-serine/threonine kinase called Akt.
•PIP3 binds to a domain of Akt known as the pleckstrin homology
domain. This interaction recruits Act to the inner face of the plasma
membrane, where it is phosphorylated and activated by other protein
kinases (called PDKs) that also contain pleckstrin homology domains
and bind PIP3.
•The formation of PIP3 thus results in the association of both Akt and
PDKs with the plasma membrane, leading to phosphorylation and
activation of Akt. Once activated, Akt phosphorylates a number of
target proteins, including proteins that are direct regulators of cell
survival, transcription factors, and other protein kinases that regulate
cell metabolism and protein synthesis.
• Second messengers can also be derived from other phospholipids.
The hydrolysis of phosphatidylcholine is stimulated by a variety of
growth factors, providing a second source of diacylglycerol, in
addition to that derived from PIP2.
• While PIP2 hydrolysis is a transient response to growth factor
stimulation, the hydrolysis of phosphatidylcholine typically persists
for several hours, providing a sustained source of diacylglycerol
that may be important in signaling long-term responses, such as
cell proliferation.
• Sphingomyelin is also cleaved in response to a variety of
extracellular stimuli, resulting in the formation of ceramide.
Although its targets remain to be fully elucidated, ceramide
regulates a number of protein kinases and phosphatases that can
affect cell proliferation and survival.
SUMMERY RELATED TO IP3 / DAG :
 IP3 /DAG PATHWAY FOLLOWED BY :

• Epinephrine (α1)
• Acetylcholine(M1 , M3)
• Angiotension
• GnRH (Gonadotropin Releasing Hormone)
• GHRH (Growth Hormone Releasing Hormone)
• Oxytocin
• TSH (Thyroid stimulating hormone)
 REFERENCE :

• http.//wikipedia.org
• Books - Goodman & Gillman
- M.N. Ghosh
- K D Tripathi.
- Rang & dale’s
THANK YOU