SGD 3: SIGNAL TRANSDUCTION

Presentation: Aug 25 (Wed)

Leader: (1) GEN

1. Discuss the various interactions between ligands and receptors (5)- Angge, Jesli, Isa, Serna, Josh
a. Affinity
b. Competition
c. Saturation
d. Specificity
2. Compare and contrast the signal transduction pathway between tyrosine kinase receptors and steroid hormone receptors
(3) - Ann, Clare, Carlo
3. Illustrate the G protein signal transduction pathway involved in the pathophysiology of Vibrio cholera associated diarrhea. (3)
Karelle, Paula, Azi

● Discuss the various interactions between ligands and receptors (Affinity, Competition , Saturation , Specificity) (5)

● Ligand-Receptor Binding - josh

○ All signaling molecules bind to specific receptors that act as signal transducers, thereby converting a ligand-receptor binding
event into intracellular signals that affect cellular functions.

● Affinity - Serna
○ The degree to which a ligand binds to a target receptor.
○ Ligands with low affinity can be displaced from a target receptor by ligands with higher affinity for the same receptor.
○ It is a function of both the rate of association and the rate of dissociation of the ligand-receptor complex
■ Ligands depend on the specificity at a molecular level
■ Receptors depend on the strength of the chemical bond where the ligand attaches
○ Systems requiring fine modulation must have agonists with low receptor affinity because with high receptor affinity would
produce unnecessarily prolonged responses.
■ Nerve synapses
○ During stimulation, agonist concentration near the receptor must be relatively high to achieve effects at concentrations that are
difficult in vivo
■ Growth factors, peptides with very high affinity with their receptors
○ Some drug-receptor interactions are so strong that they are irreversible
■ Aspirin, irreversibly inhibits its target, the enzyme cyclooxygenase

● Competition - Jesli
○ This is the interaction of different molecules that are structurally the same to compete with each other for the same receptor.
Different ligands can have the same binding site.
○ If two competing ligands, A and B, are present, increasing the concentration of A will increase the amount of A that is bound,
thereby decreasing the number of sites available to B and decreasing the amount of B that is bound.
■ So a take away from this simple example is that a biological effect of a ligand can be diminished by the
presence of another; and again if you increase the amount of that substance it will decrease the binding sites
for the other.
○ To further explain this, here is an example of a drug that would compete for a receptor. *picture*
■ Here is a simple picture where propranolol competes with your catecholamines(norepinephrine and
epinephrine)
● Agonist and Antagonist - Josh
○ Agonist - a chemical messenger which binds to the receptor and triggers the cell response. It may be in a form of hormones,
neurotransmitters such as acetylcholine, histamine, norepinephrine or in a form of drug
 ○ Antagonist - is a molecule that competes with the receptor with a chemical messenger that is normally present in the body. It
binds to the receptor but there would be no response.
● Upregulation
○ Upregulation is the increase of the number of receptors when the activity of the receptor is lower than usual. When the level of
a hormone is chronically reduced, target cells engage in upregulation to increase their number of receptors. This process
allows cells to be more sensitive to the hormone that is present. Cells can also alter the sensitivity of the receptors themselves
to various hormones.
● Down regulation
○ Down regulation is the decrease of the number of receptors when the activity of the receptor is higher than usual due to
repeated exposure. It allows cells to become less reactive to the excessive hormone levels.
● Saturation - Isa
○ Percentage of the total number of specific receptor that is occupied by messengers
○ Fraction of total binding sites that are occupied at any given time
■ A receptor has a limited number of binding sites → Saturated at high ligand concentrations
● For example, if all receptors are occupied, they are fully saturated or 100% saturated; if half are
occupied, the saturation would be 50%
● A single binding site would be 50% saturated if it were occupied by a ligand 50% of the time
○ Factors affecting the percent saturation of a binding site
■ Concentration of unbound ligand in the solution
● Percent saturation of binding sites increases with increasing ligand concentration until all the sites
become occupied
● Assuming that the ligand is a molecule that exerts a biological effect when it binds to a protein, the
magnitude of the effect would also increase with increasing numbers of bound ligands until all the
binding sites are occupied
● Increases in ligand concentration would produce no further effect because there would be no additional
sites to be occupied
● A continuous increase in the magnitude of a chemical stimulus (ligand concentration) that exerts its
effects by binding to proteins will produce an increased biological response until the point at which the
protein-binding sites are 100% saturated
■ Affinity of the binding site for the ligand (Discussed by Serna)
● Collisions between molecules in a solution and a protein containing a bound ligand can dislodge a
loosely bound ligand
● If a binding site has a high affinity for a ligand, even a reduced ligand concentration will result in a
high degree of saturation because, once bound to the site, the ligand is not easily dislodged
● A low-affinity site, on the other hand, requires a higher concentration of ligand to achieve the same
degree of saturation
● One measure of binding site affinity is the ligand concentration necessary to produce 50% saturation;
the lower the ligand concentration required to bind to half the binding sites, the greater the affinity of the
binding site
● Specificity- Angge
○ Ability of the receptor to bind exclusively to one type or a limited number of structurally related types of chemical messenger.
○ If a chemical messenger is released in the bloodstream, it will bind to the cells which only contain its specific receptor.
○ Three of the most common ligands that are distributed in the body are acetylcholine, epinephrine and norepinephrine.
○ Cholinergic Receptors
■ Acetylcholine receptors are known as cholinergic receptors.
■ It is further sub-classified as nicotinic and muscarinic. Each has a specific subtype which is summarized as
follows:
○ Adrenergic Receptors
■ Epinephrine and norepinephrine receptors are known as adrenergic receptors. It is further sub-classified as
alpha and beta receptors.
■ Each has a specific subtypes which is summarized in this table

○ A cell can contain numerous types of receptors.

■ If acetylcholine is released into the circulation, it is going to bind to the cell's cholinergic receptor but not to the
adrenergic receptor. The cholinergic receptor is specific only for the acetylcholine.
○ There are also instances wherein an organ has 2 receptor Types for a Single Ligand
■ A vascular smooth can contain both alpha 2 and beta 2 receptors for epinephrine.
■ The response of the cell may be different for the same ligand
■ This is based on the attached type of G protein (G stimulatory & G inhibitory)
● GS- relaxation
● GI- contraction
 ■ Stimulation
● Binding of epinephrine to beta 2 receptors activates GS protein complex which stimulates adenylyl
cyclase that leads to INC cAMP
● cAMP activates PKA, leading to intestinal smooth muscle RELAXATION
● Beta 2 → vascular smooth RELAXATION
■ Inhibition
● Binding of epinephrine to alpha 2 receptors activates GI protein complex which does not result to
activation of adenylyl cyclase
● Leads to reduction in the levels of cAMP and PKA, resulting to smooth muscle CONTRACTION
● Alpha 2 → vascular smooth CONTRACTION
○ Cross Talk
■ Due to the presence of different signal transduction pathways within a cell, some pathways induce exactly
opposite cellular response
■ An example of this is the cross talk between B2 and M3 receptors
■ B2 activation by epi as part of sympathetic response→ BRONCHODILATION
■ M3 activation by acetylcholine as part of the parasympathetic response → BRONCHOCONSTRICTION

Supplementary
● Agonist vs Antagonist
● Down-regulation and Up-Regulation

● Compare and contrast the signal transduction pathway between tyrosine kinase receptors and steroid
hormone receptors. (Ann, Clare, Carlo)

● Tyrosine kinase Receptors (Ann)

○ Definition
○ Signal transduction pathway
■ There are two classes of TKR
■ Nerve growth factor (NGF) receptors are typical examples of one class
■ Ligand binding to two NGF receptors facilitates their dimerization → phosphorylation of cytoplasmic tyrosine
kinase domain of each monomer to phosphorylate and activate the other monomer
■ Once the other monomer is phosphorylated, the cytoplasmic domains can recruit GEFs such as growth
factor-receptor bound protein 2 to the plasma membrane which in turn activates Ras and downstream kinases
that regulate gene transcription programs important for cell survival and proliferation
○ Example
■ Activation of the insulin receptor (which is tetrameric and composed of 2 alpha and 2 beta subunits) by insulin
is an example of the other type of tyrosine kinase receptor
■ Binding of insulin to the alpha subunits produces a conformational change that facilitates interaction between
alpha and beta pairs
■ Binding of insulin to its receptor causes autophosphorylation of tyrosine residues in the catalytic domains of the
beta subunits and activated receptor then phosphorylates cytoplasmic proteins to initiate its cellular effects
● Including stimulating the absorption of glucose from the blood into skeletal muscle and fat tissue

● Steroid hormone receptors (Clare)

STEROID HORMONE RECEPTORS [CLARE]
○ Definition
■ Steroid hormone receptors belong to the class of nuclear receptors, in which there is the presence of small
hydrophobic molecules that have a long biological half-life and can diffuse across the plasma membrane.
■ The steroid hormone may first bind to either a nuclear receptor or to a cytoplasmic receptor.
■ And once bound, the goal would be to translocate to the nucleus where transcription of specific genes will
occur.
○ Signal transduction pathway & Example [Figure]
■ For example, the hormone aldosterone which is a mineralocorticoid may diffuse across the plasma membrane
because of its hydrophobic characteristic.
■ It will then interact with a chaperone such as the heat-shock protein, found together with the nuclear receptor
in a complex. This will cause a conformational change that will dissociate the heat-shock protein from the
receptor, and facilitate the translocation of the hormone-bound receptor complex to the nucleus.
■ In the nucleus, the receptor also binds to a coactivator protein that will activate gene transcription. Once
activated, the nuclear receptor will then bind to a specific DNA sequence called the hormone response
element and transcription will be initiated leading to several processes that can change cell function.
■ *Sources: Berne, https://opentextbc.ca/biology/chapter/18-2-how-hormones-work/
● Similarities of Tyrosine kinase and steroid hormone receptors (Carlo)
○ Both the Tyrosine kinase and Steroid hormone receptors pathways need an intracellular cytoplasmic coupling with
proteins to be able to reach the target gene. In the Tyrosine Kinase pathway, the protein would be the STAT or signal
transducer and activator of transcription proteins. This in turn will create a downward cascade of repeated
phosphorylation as it will reach the target gene and initiate transcription.
 ○ On the other hand, for the steroid hormone pathway, a heat shock protein, which functions as a chaperone will assist
in the delivery of the hormone-receptor complex to the nucleus. This will further initiate or inhibit gene transcription.
○ Both pathways end product is the synthesis of protein by activation of gene transcription and formation of mRNA
Source: Guyton

● Illustrate the G protein signal transduction pathway involved in the pathophysiology of Vibrio cholera
associated diarrhea. (3)

● Definition of Terms
○ Vibrio Cholerae
■ disease produced: cholera
■ Vibrio cholerae is a species of Gram-negative, facultative anaerobe and comma-shaped bacteria.
■ MOT: Contaminated water thru oral fecal route
■ Habitat: Intestinal Lumen and infects intestinal epithelial cells
○ Signs and Symptoms
■ Gastroenteritis, Severe Diarrhea
● Causing a loss of 5 to 10 liters of water and sodium chloride as diarrhea each day
■ Caused by: AB enterotoxin
● 2 subunits
○ 5 B subunits - can bind to host cell receptor (GM1)
○ 1 A subunit with 2 domains
■ A1 - active & A2
Note: The intestinal epithelial cells contain chained GM1 ganglioside receptors which binds to the cholera toxin, which allow the AB
toxins from the cholera to internalize.

○ G protein and Signal Transduction

■ 3 Subunits:
● Alpha - binds and hydrolyzes GTP and interacts with effector proteins; anchors GPCR complex to the
membrane
○ Inactive - carries GDP
■ GPCR (G protein Coupled Receptor) has a GDP attached to its alpha subunit and it
remains as a single protein (alpha, beta, and gamma subunits are all attached to each
other)
○ Active - carries GTP
■ Can bind with Adenylate cyclase;
● adenylate cyclase can convert ATP to cAMP
■ GDP is replaced with GTP wherein it dissociates into 2 components:
● (1) alpha and (2)beta gamma.
● Upon receptor activation, the G protein undergoes conformational change.
■ When active, the active Alpha subunit detaches, and binds to activate adenylate
cyclase, ultimately producing cAMP as the GTP goes back to inactivated GDP --
producing only small amounts of GDP

● Beta - interacts with effector proteins distinct from that of the alpha subunit
● Gamma - interacts with effector proteins distinct from that of the alpha subunit; anchors GPCR complex
to the membrane
PATHOPHY
Simplified pathophysiology (Berne and Levy) AZI
● Vibrio cholerae releases cholera toxin
○ Cholera toxin binds to the GM1 ganglioside receptor
○ Toxin will go to endoplasmic reticulum
○ A1 subunit of the enterotoxin is released into the cytoplasm
● Receptor activation
○ GDP is replaced by GTP and G protein dissociates into two components:
■ Alpha subunit which is attached to the GTP
■ Beta and gamma subunits
● Alpha 1 subunit of the cholera toxin binds to the G protein using NAD+ which it will use to bind with the G protein.
○ NAD contains Nicotinamide, a Ribose sugar, and ADP
● Toxin serves as a catalyst in the ADP ribosylation of the α subunit of the G-protein (s-type)
○ NAD binds to G protein
○ Dissociate into:
■ Nicotinamide portion
■ Ribose-adp will remain bound to the g protein

Karelle

● Result: inhibits GTPase activity of 𝑎 subunit

○ it prevents G protein from cleaving GTP into GDP
● 𝑎 subunit remains in GTP bound state which is its activated configuration
● ↑ adenylyl cyclase remains active for a long period
● ↑ thus increasing the levels of cyclic AMP tremendously
○ Elevated levels of cAMP stimulates a host enzyme which is Protein Kinase A (enzymatic activity activates various iron
transport channels)
● ↑ Protein Kinase A activation
● Organ affected: intestines
○ ↑ PKA ↑ cystic fibrosis transmembrane conductance regulator (CFTR) -responsible for the export of Chloride
○ ↑ efflux of chloride ions from inside the cell into the interstitial crypts
● “As chloride and other ions leave the cell, water leaves as well.”
○ Because of the attempt to equilibrate the osmolarity, the bloodstream provides water chloride and ions --->leads to the
activation of sodium pumps
○ All this extra sodium chloride causes extreme osmosis of water from the blood, thus providing rapid flow of fluid along
with the salt.
● Result in: Secretory Diarrhea
○ Hallmark presentation of cholera
○ Appears profuse and watery
● Extensive loss of fluids
○ Could cause dehydration

------END-----
○
The relationship of Vibrio Cholerae with the G Protein Signal Transduction Pathway
● The cholera toxin contains 2 subunits which A (1) and B (5) subunits and it affects G-protein signalling as it’s responsible for
causing a covalent and irreversible activation of the G protein, which in response, simply activates the adenylate cyclase and
raises intracellular cAMP levels located at the basolateral membrane of the intestinal epithelial cells. When the cyclic-AMP
levels increase, it also alters the transport of Na+ and Cl-, through the activation of the CFTR and which would result in a huge
amount of chloride loss.
● When chloride ion is lost in the intestinal lumen, water goes inside the lumen and causes large amounts of diarrhea and fluid
loss from the body

DRAFT MOA:
○ The A1 od the Cholera toxin stimulates formation of excess cyclic adenosine monophosphate, which opens
tremendous numbers of chloride channels, allowing chloride ions to flow rapidly from inside the cell into the intestinal
crypts.---------> activate sodium pump
○ All this extra sodium chloride causes extreme osmosis of water from the blood, thus providing rapid flow of fluid along
with the salt.
○ When the A1 subunit of the enterotoxin binds to the G protein, it uses an NAD+ which it will use to bind with the G
protein. Then, the nicotinamide from the NAD will be left after binding to the G Protein; what is now binded is the
ribose-adp-G protein unit. Also known as an ADP ribosylation reaction.
○ Ribosylation reaction
■ Prevents the GTP from going back to a GDP, hence the cAMP formation will not be halted. This produces
HUGE amounts of cAMP in the cell which stimulates a reaction that will increase CHLORIDE ions to go out of
the cell. Simultaneously, letting water out of the cell to equilibrate the osmolarity
■ Thereby, the blood stream compensates by producing more chloride and water to the area which ultimately
causes diarrhea which helps the spread of the bacteria.

■ Pathogenesis:
● Excessive secretion of electrolytes
● Cholera Toxin (enterotoxin)
○ B subunit (5 copies) - binds to host cell receptors
○ A subunit (1 copy) - Exhibits the toxic activity
■ A1 domain - binds to the G protein

● Summary (Leader)
To summarize all of the points that were raised by each of the discussants wherein we will be starting with the
● Various Interactions Between Ligands and Receptors
○ So, we have your ligand-receptor binding that will be converted or acting as the signal transducers and will affect
different cellular functions.
○ For the interactions,
■ So, we discussed first the AFFINITY, it is the degree to which a ligand binds to a target receptor.
● Ligands with low affinity can be displaced from a target receptor by ligands with higher affinity for the
same receptor.
■ The second would be COMPETITION. This is the ability of different molecules that are structurally the same to
compete with each other for the same receptor.
■ The third would be SATURATION. This is going to dictate the percentage of the total number of specific
receptors that is occupied by messengers.
■ And lastly, SPECIFICITY. This is the Ability of the receptor to bind exclusively to one type or a limited number of
structurally related types of chemical messenger.

● Signal transduction pathway between

○ tyrosine kinase receptors and steroid hormone receptors
■ Basically, they would both be needed in intercellular cytoplasmic coupling with proteins, activation of gene
transcriptions and formation of mRNA
■ In the tyrosine kinase pathway
● the response element would be the STAT or signal transducer and activator of transcription
proteins while the
■ steroid hormone receptor pathway, the response element will be the receptor itself or can be found in the
nucleus where coupling with the hormone-receptor complex will initiate or inhibit gene transcription.

● G protein signal transduction pathway and vibrio chordae

■ This will have three different subunits which will stimulate the activation of the sodium pump
■ In relation to vibroe chordae, this will release a toxin called cholera
■ And this cholera will be eliciting an irreversible activation of the G-protein signalling
■ And then activating the adenylate cyclase
■ Which will be further prompt the loss of chloride ions leading to diarrhea and loss of fluid inside the body.

● References