DR - Imran Ali: (AP Biochemistry)

Dr.
Imran Ali
(AP Biochemistry)
Dr.Imran Ali Biochem

 Learning objectives

 At the end of this session, students will be able to
define explain and interpret
 G proteins
 Receptors and target cells
 G protein coupled receptors(GPCRs)
 Ist messenger/2nd messenger
 Cell signaling pathways

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Agonists versus antagonists
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 Not every ligand that binds to a receptor also activates that receptor.
The following classes of ligands exist:
 Agonists are able to activate the receptor and result in a strong
biological response.
 Partial agonists do not activate receptors with maximal efficacy, even
with maximal binding, causing partial responses compared to those
of full agonists
 Antagonists bind to receptors but do not activate them. This results in
a receptor blockade, inhibiting the binding of agonists and inverse
agonists
 Allosteric modulators: They do not bind to the agonist-binding site of
the receptor but instead on specific allosteric binding sites, through
which they modify the effect of the agonist.
RECEPTOR
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Adrenergic VS Cholinergic Receptors
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Ligand-gated ion channels
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 also commonly referred to as ionotropic receptors,
are a group of transmembrane ion-channel proteins
which open to allow ions such as Na+, K+, Ca2+,
and/or Cl− to pass through the membrane in
response to the binding of a chemical messenger (i.e.
a ligand), such as a neurotransmitter

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G Protein
G Protein Coupled
Coupled
Receptors(GPCRs)
Receptors(GPCRs)

GPCRs
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GPCRs have various types
of ligands:
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GPCRs
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EXAMPLES OF GPCRs
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EXAMPLES OF GPCRs

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G-Proteins
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G PROTEINS:

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GDP
Adenyl
GTP
cyclase

Adenylyl Cyclase
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1 st messenger/2 nd
messenger

 The ligand that binds to cell membrane
receptor of target cell is called as 1st
messenger
 2nd messengers are molecules that
relay signals from cell surface receptors
to target molecules inside cells

1 st messenger/2 nd
messenger

 The ligand (1st messenger) binds to cell
membrane receptor of target cell.
 This leads to formation of second messenger
inside the cell after a series of events
 Some examples of 2nd messenger are:
 cAMP
 IP3 and DAG
 Calcium-calmodulin

GTP
GDP
Dr.Imran Ali Biochem Phosphorylates

proteins
Protein Kinase
 In Prokaryotic cells, cAMP binds to a specific protein called

catabolite regulatory protein (CRP) that binds directly to
DNA and influences gene expression.
 In Eukaryotic cells, cAMP binds to a protein kinase called
protein kinase A (PKA), a heterotetrameric molecule
consisting of two regulatory subunits (R) and two catalytic
subunits (C). cAMP binding results in the following reaction:
The R2C2 complex has no enzymatic activity, but the binding

of cAMP by R induces dissociation of the R-C complex,
thereby activating the latter

 The Active C subunit catalyzes the transfer of the
phosphate of ATP to a serine or threonine residue in
a variety of proteins which leads to a variety of
physiological cellular responses.

MOST IMPORTANT PROTEIN
KINASES

Protein kinase A………………..Through cyclic
AMP
Protein kinase B………………..E.G., insulin
tyrosine kinase mechanism
Protein kinase C………………..Through DAG
and calcium

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Phosphorylation
of protein

 i. Any ligand e.g., a hormone binds with the receptor in the cell membrane
and forms the hormone-receptor complex
 (ii) It activates the G protein.

 iii. G protein releases GDP from α-GDP unit
 iv. α-subunit now binds with new molecule of GTP, i.e. the GDP is
exchanged for GTP
 v. This exchange triggers the dissociation of α-GTP unit and β-γ dimmer
from the receptor
 vi. The α-GTP unit activates the enzyme adenyl cyclase(AC), which is
present in the cell membrane. Most of the adenyl cyclase protrudes into the
cytoplasm of the cell from inner surface of the cell membrane
 vii. Activated AC converts the adenosine triphosphate of the cytoplasm
into cyclic adenosine monophosphate (cAMP 2nd messenger) which
activates protein kinase A Dr.Imran Ali Biochem
 Viii. Phosphorylation of proteins:
 Activation of protein kinase A causes addition of a phosphate group to a
protein leads to cellular response
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• Smooth muscle contraction of

blood vessels, eye, prostate, bladder
Dr.Imran Ali Biochem • Salivary gland secretion
• Gastric acid secretion
• CNS Excitation
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NERVOUS SYSTEM?

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Gq Gi

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Dephosphorylation of
proteins:
added to proteins by
The phosphate groups
protein kinases are removed by protein
phosphatases
 —Enzymes that hydrolytically cleave

phosphate esters. This ensures that changes
in protein activity induced by
phosphorylation are not permanent.
Phosphodiesterases

GTPases

The actions of the Gα–GTP complex are
short-lived because Gα has an inherent GTPase activity.
GTPases (singular GTPase) are a large family of hydrolase

enzymes that can bind and hydrolyze guanosine triphosphate
(GTP).

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CHOLERA
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Primary pathways of cell signaling
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G-Protein mediated pathway:
Adenylate cyclase pathway
Phospholipase mediated pathway
Non-G protein mediated pathway:
Intracellular receptor mediated pathway(HRE)
Receptor tyrosine kinase mediated pathway

JAK/STAT pathway
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Phosphatidylinositol and diacylglycerol
: 2nd messenger system
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Phosphorylation
of protein
Phospholipase C
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γ
β γ
OXYTOCIN
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I. Certain hormone–receptor interactions result in the
activation of PLC. This appears to involve a specific
G protein, which also may activate a calcium
channel. PLC results in generation of IP3 which
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liberates stored intracellular Ca2+, and DAG a
potent activator of protein kinase (PKC). In this
scheme, the activated PKC phosphorylates specific
substrates, which then alter physiologic processes.
Likewise, the Ca2+-calmodulin complex can
activate specific kinases.
II. These actions result in phosphorylation of
substrates, and this leads to altered physiologic
responses.
III. Ca2+can enter cells through voltage- or ligand-
gated Ca2+channels. The intracellular Ca2+ is also
regulated through storage and release by the
mitochondria and endoplasmic reticulum.
INSULIN:

SOURCE OF SECRETION:
• Insulin is secreted by the β-cells in the islets
of Langerhans of pancreas.
• Insulin is only hormone in the body that
reduces blood glucose levels.
INSULIN RECEPTORS
 Insulin binding to its receptor stimulates
intrinsic tyrosine kinase activity leading to
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receptor auto phosphorylation and the
recruitment of intracellular signaling
molecules, such as insulin receptor substrates
(IRS).
 IRS and other adaptor proteins initiate a
complex cascade of phosphorylation and

dephosphorylation reactions, resulting in the
widespread metabolic and mitogenic effects of
insulin.

INSULIN RECEPTOR STRUCTURE
(For insulin binding) 
(For ATP binding)

. The receptor is composed of two extracellular α-subunits
that are each linked to a ß-subunit and to each other by disulfide
bonds.
 /PKB

GRB2 (Synthesis of lipids
Proteins and glycogen)
(Cell growth, proliferation,
gene expression)
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MCQ PRACTICE:
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A 12-year-old type 1 diabetic female patient checks her
blood glucose through a glucometer before taking meal.
She then injects herself subcutaneously with exogenous
insulin and then takes her meal. As Insulin is absorbed in
to her blood, it binds to insulin receptors that activates:
A. Tyrosine kinase
B. Adenylate cyclase
C. Cyclic AMP
D. Protein kinase C
E. Phospholipase C
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Guanylyl cyclase-cGMP
2nd messenger system

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Ligands for cGMP
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Examples are:
Nitric oxide
Atrial Natriuretic peptide(ANP)

Atrial natriuretic peptide
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 The main function of ANP is causing a reduction in
expanded extracellular fluid (ECF) volume by
increasing renal sodium excretion.
 ANP is synthesized and secreted by cardiac muscle
cells in the walls of the atria in the heart.
 These cells contain volume receptors which respond
to increased stretching of the atrial wall due to
increased atrial blood volume.

Atrial natriuretic peptide
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 Reduction of blood volume by ANP can result in
secondary effects such as reduction of extracellular
fluid (ECF) volume (edema), improved
cardiac ejection fraction with resultant improved
organ perfusion, decreased blood pressure,
JAK-STAT Signaling
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MCQ PRACTICE
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NF- κB Pathway
(NF- ΚB is basically a transcription factor)

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CREB
 The transcription factor NF- κB is a heterodimeric complex
typically composed of two subunits termed p50 and p65
 Normally, NF- κB is kept sequestered in the cytoplasm in a
transcriptionally inactive form by members of the Inhibitor
of NF- κB (IκB) family. 
 Extracellular stimuli such as proinflammatory cytokines,
reactive oxygen species, and mitogens lead to activation of
the IκB kinase complex, IKK, which is a heterohexameric
structure consisting of α, β, and γ subunits.
 IKK phosphorylates IκB on two serine residues, and this
targets IκB for ubiquitination and subsequent degradation by
the proteasome.
 Following IκB degradation, free and activated NF-
κB(P50+P65) translocates to the nucleus, where it binds to a
number of gene promoters such as CREB and activates
transcription, particularly of genes involved in the
inflammatory response.

NF- κB Pathway inhibitors:
Glucocorticiods

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Hormone response
element (HRE)

 A hormone response element (HRE) is a short
sequence of DNA within the promoter of a gene that
is able to bind to a specific hormone receptor
complex and therefore regulate transcription
 In genetics, a promoter is a region of DNA that leads

to initiation of transcription of a particular gene.



 Thank you

DR - Imran Ali: (AP Biochemistry)

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Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem Phosphorylates

Dr.Imran Ali Biochem

The R2C2 complex has no enzymatic activity, but the binding

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

• Smooth muscle contraction of

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

 —Enzymes that hydrolytically cleave

Dr.Imran Ali Biochem

GTPases (singular GTPase) are a large family of hydrolase

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem

Dr.Imran Ali Biochem