Principles of Signal Transduction: Objectives

1. Be able to compare and contrast the overall properties of

endocrine and synaptic signaling mechanisms.
a. Endocrine Signaling Mechanism:
i. Hormone is secreted by an endocrine gland and is
carried, via the bloodstream, to a distant target cell
ii. Signaling is SLOW
1. Relies on passive diffusion and blood flow
iii. Signals are DILUTE
1. Receptors have high specificity and affinity for
the signal
a. Chemically based specificity and affinity
2. There is a slow termination of the response
(slow dissociation rate)
iv. Endocrine cells secrete many ligands thus the
target cells must be specific with a high affinity
v. Example: Steroids
b. Synaptic Signaling Mechanism:
i. Signaling is FAST
1. Uses electrical impulses over short distance
ii. Signals reach high local concentrations
1. Receptors have low affinity for the signal
a. Mechanically based (based on the
placement of the synapses)
2. There is a rapid termination of the response
(quick dissociation rate)
iii. Secrete few distinct ligands the specificity comes
from the precise nerve/target cell contact point
iv. Examples: Peptides and proteins; catecholamines

Principles of Signal Transduction: Objectives

2. Be able to identify the four major types of signal transduction
pathways and give examples of ligands and cellular responses
for each.
a. Receptor Tyrosine Kinase
i. Ligands: Insulin, lots of growth factors
ii. Insulin Receptor Signaling:
1. Receptor binds two signal compounds (in this
case, insulin peptide) which activates a
tyrosine autokinase. This also phosphorylates
IRS-1 (Insulin Receptor Substrate 1).
iii. IRS-1 has three tissue specific pathways
1. Growth: IRS1 phosphorylates Shp, activating
Ras.
a. Ras activates MAP which activates
Transcription factors which promote
growth
2. Glucose Uptake
a. IRS-1 phosphorylates P13K, initiating a
phosphorylation cascade
i. Leads to GLUT4 (transporters of
glucose) onto the plasma
membrane
3. Glycogen Deposistion
a. IRS1 phosphorylates P13K, initiating a
phosporylation cascade
i. Leads to stimulation of enzymatic
steps in the conversion of glucose
to glycogen
b. G Protein-Linked Receptors
i. Ligands: a great variety use G Protein-Linked
Receptors
1. Catecholamines, NTs, Peptide hormones
ii. 2 possible cellular responses:
1. cAMP Signal Pathway
a. cAMP activates cAMP-dependent Protein
Kinase (PKA) by phosphorylating PKA
i. Kinase is a tetramer
1. 2 cAMP-binding chains
2. 2 catalytic chains
b. Binding causes release of activated
catalytic subunits
c. Catalytic units phosphorylate substrates
(usually enzymes)
d. Moderation or reversal of response is
achieved via:
i. Dephosphorylation of substrates
1

Principles of Signal Transduction: Objectives

1. By phosphatases
ii. Degradation of cAMP
1. By phosphodiesterases (PDE)

Principles of Signal Transduction: Objectives

2. Phosphatidylinositol Pathway
a. Hormone signal causes increase in
cytosolic Ca++ and activation of PKC,
leading to 2 distinct (but interacting)
chain of events:
i. Ca++ binds to Calmodulin (CaM)
1. Myosin Light Chain Kinase
(MLCK) phosphorylates
myosin
a. Causes interaction with
actin and muscle
contracts
2. Nitric Oxide Synthase (NOS)
ii. Protein Kinase C phosphorylates a
variety of enzymes and proteins
1. Ion channels (like Na+/H+
pump)
a. Phosphorylation leads
to increase of cellular
pH and proliferation
2. Activation of transcription
factors controlling gene
expression
c. Intracellular Receptors (steroids)
i. Examples of ligands: Glucocorticoid, mineralcorticoid,
progesterone, estrogen, androgen, vitamin D, thyroid
ii. Three domains
1. Transcription activation Domain
a. Responsible for interaction promoting
activity by RNA polymerase
i. Thus, Communication with Pol
2. DNA Binding Domain
a. Site of direct interaction with DNA in the
promoter region of genes
i. Thus, Recognition of Hormone
Response Elements
3. Ligand Binding Domain.
a. Site of high-selectivity binding by steroid
hormones
i. Thus, Binding Site
iii. 2 Activated Steroid Receptors recruit the Histone
Acetyl Transferase (HAT) Complex
1. Leading to acetylation of histones
a. Causes DNA to unwind
i. Allowing the binding of the
transcription apparatus.
1

Principles of Signal Transduction: Objectives

Principles of Signal Transduction: Objectives

d. Ligand Gated Channels
i. Converts extracellular signals into electrical impulses
ii. Occurs between nerves and target cells (Chemical
Synapse)
1. Nerve terminal releases neurotransmitters
(Ach, GABA, etc) by fusion with storage
vesicles with plasma membrane
2. NT binds channels present in synapse region of
target cell
3. Gated channel opens, allowing entry of ions
iii. Example: Nicotinic Acetylchnoline Receptor:
1. Channel opening requires 2 molecules of ligand
2. Channel opens only briefly
a. Closes while still ligated
3. Ligand dissociates, returning channel to resting
state
4. Dissociated ACh is hydrolyzed by
cholineresterase
3. Be able to identify or describe the key steps and molecules
involved in each of the four major pathways, including structure
and location of receptors, 2nd messenger molecules (including
calcium and nitric oxide), effector enzymes, and substrates.
a. Ligand-Gated Channels
i. Key Steps:
1. Nerve terminal releases neurotransmitters
a. Via fusion of storage vesicles with plasma
membrane
2. NT binds channels present in synapse region
3. Gated channel opens, allowing entry of ions
ii. Molecules involved:
1. Neurotransmitters
a. ACh, GABA, Serotonin, Glutamate,
Glycine
iii. Structure of receptors:
1. Composed of 5 transmembrane polypeptides
a. Each chain ~500 AAs and traverses
membrane four (4) times
2. 1 helix of each chain contains polar AAs:
Aqueous Pore
3. Negatively-charged AAs are at the mouth of
the channel
a. Prevents entry of anions (ligand
specificity)
iv. Location of receptors:
1. Plasma membrane of Target Cells
(transmembrane)
1

Principles of Signal Transduction: Objectives

v. 2nd messenger molecules:
vi. Effector Enzymes:
1. Cholineresterase (hydrolyzes dissociated ACh)
vii. Substrates:

Principles of Signal Transduction: Objectives

b. Receptor Tyrosine Kinases:
i. Key Steps:
1. Ligand binding to the insulin receptor (IR)
a. Induces phosphorylation on tyrosine
residues on the intracellular domain
b. 2 ligands of insulin peptide are required
for full activation
ii. Molecules involved:
1. Insulin Peptide
2. IRS1
3. Ras (for MAP kinase cascade to promote growth
factors)
4. P13K (is phosphorylated by IRS1, initiates
phosphorylation cascade)
iii. Structure of receptors:
1. Single transmembrane alpha helix
2. Example: Insulin
a. 2 beta and 2 alpha chains (sulfide bond
connects them)
b. Alpha chains: contain hormone-binding
site
c. Beta chains: traverse the membrane and
contain tyrosine kinase domain
iv. Location of receptors:
1. On the plasma membrane (transmembrane)
2. Extracellular domain
a. Ligand-binding site
3. Intracellular domain
a. Tyrosine kinase activity
v. 2nd messenger molecules:
1. IRS1
2. Ras(for MAP kinase cascade to promote growth
factors)
3. P13K (is phosphorylated by IRS1, initiates
phosphorylation cascade)
vi. Effector Enzymes:
1. MAP Kinase (activates transcription factors to
promote growth)
vii. Substrates:

Principles of Signal Transduction: Objectives

c. Steroid (Intercellular) Receptors:
i. Key Steps:
1. 2 Ligand-ed Steroid Receptors recruits a coactivator that contains the enzyme, Histone
Acetyl Transferase (HAT)
2. That enzyme leads to acetylation of histones
3. The acetylated histones allow the DNA to
unwind
4. The unwound DNA allows for the binding of
general transcription factors (such as Pol II)
ii. Molecules involved:
1. SR (Steroid Receptor)
2. HSP
3. IP
4. CoA Complex
5. TATA (gene expression begins here via RNA
transcription)
iii. Structure of receptors:
1. NH2 Transcription activation domain DNA
binding domain Ligand binding domain
COOH
2. Receptors are always ON
iv. Location of receptors:
1. Plasma membrane
2. Cytosol
3. Nucleus
nd
v. 2 messenger molecules:
vi. Effector Enzymes:
1. Histone Acetyl Transferase (HAT)
a. Acetylates histones
b. Leads to the unwinding of DNA
c. Allows for binding of general
transcription factors
vii. Substrates:

Principles of Signal Transduction: Objectives

d. G-Protein Linked Receptors
i. Structure of receptors:
1. 7 Pass transmembrane receptor
a. Generates 4 intracellular and
extracellular loops
b. Extracellular amino terminus:
i. Ligand binding site
c. Intracellular loops:
i. G Protein Interaction
ii. Phosphorylation-mediated
inactivation
1. When 3rd loop is
phosphorylated, the receptor
is inactivated
ii. cAMP Signaling:
1. Key Steps:
a. Hormone stimulateion causes rapid
increase in cAMP
b. Adenylate cyclase cleaves ATP to
generate cAMP and pyrophosphate
i. IRREVERSIBLE step
c. Degradation of cAMP by
phosphodiesterase
i. Yields 5-AMP

Principles of Signal Transduction: Objectives

Principles of Signal Transduction: Objectives

Principles of Signal Transduction: Objectives

2. Molecules involved:
a. cAMP, PKA (cAMP-dependent protein
kinase)
3. Location of receptors:
a. On Plasma membrane
i. Alpha chain:
1. Binds and hydrolyzes GTP
2. Activates adenylate cyclase
ii. Beta and Gamma chains:
1. Anchors alpha chain to the
cytoplasmic face
4. 2nd messenger molecules:
a. Cyclic AMP (cAMP)
5. Effector Enzymes:
a. Adenylate cyclase cleaves ATP to
generate cAMP and pyrophosphate
i. IRREVERSIBLE step
b. Degradation of cAMP by
phosphodiesterase
i. Yields 5-AMP
6. Substrates:
iii. Ca++/Phosphoinositide Signal Pathway:
1. Key Steps:

2. Molecules involved:
a. Cam (Calmodulin)
3. Location of receptors:
a. Plasma membrane
i. On they cytosolic face
4. 2nd messenger molecules:
a. Diacylglycerol (DAG)
1

Principles of Signal Transduction: Objectives

b. Inositol Trisphosphate (IP3)
c. Ca++ Ion
5. Effector Enzymes:
a. MLCK (phosphorylates myosin, causing
muscle contraction)
b. Nitric Oxide Synthase (NOS)
c. Protein Kinase C (phosphorylates lots of
enzymes and proteins)
6. Substrates:
4. Be able to describe the major mechanisms by which each signal
pathways can be inhibited.
a. Tyrosine Kinase, Steroid, and Ligand-Gated Channels:
i. Down-regulation of Cell Surface Receptors
ii. Prolonged hormone exposure leads to receptor
degradation

iii.
b. G-Linked
i. Inactivation of receptors via phosphorylation:
1. Excess hormone
a. Results in receptor that can bind
hormone but not activate G protein
2. Phosphorylation of serines and threonines on
intracellular loops
3. A-kinase first to phosphorylate when cAMP
levels get too high, followed by BARK
4. Fully phosphorylated receptor is further
blocked from G protein by beta arrestin