Cell Signaling

PowerPoint® Lecture Presentations for by

Biology Soner Dogan, PhD

Eighth EditionProfessor of Medical Biology
Neil Campbell and Jane Reece
Yeditepe University
School
Lectures by Chris Romero, updated of Barley
by Erin Medicine
with contributions from Joan Sharp
Copyright © 2008 Pearson Education, Inc., publishing as Pearson Benjamin Cummings .
 nowdona
whathari

How do the effects of Viagra (multicolored) result from its inhibition of a

signaling-pathway enzyme (purple)?
 Intercellular Junctions

• Neighboring cells in tissues, organs, or organ

systems often adhere, interact, and communicate
through direct physical contact

• Intercellular junctions facilitate this contact

• There are several types of intercellular junctions
– Plasmodesmata (in plant cells)
– Tight junctions
– Desmosomes in animal cells

– Gap junctions
 signal transduction

• A signal transduction pathway is a series of

steps by which a signal on a cell’s surface is
converted into a specific cellular response
 Signaling Molecules and Their Receptors
• Endocrine signaling—signaling molecules (hormones)
are secreted by specialized endocrine cells and carried
through the circulation to target cells at distant body
sites. Example: estrogen.
• Paracrine signaling—molecules released by one cell act
on neighboring target cells,
e.g., neurotransmitters.
• Autocrine signaling—cells respond to signaling
molecules that they themselves produce.
Example: T lymphocytes respond to antigens by
making a growth factor that drives their own
proliferation, thereby amplifying the immune
response.
 Local and Long-Distance Signaling

• Cells in a multicellular organism communicate by

chemical messengers
• Animal and plant cells have cell junctions that
directly connect the cytoplasm of adjacent cells
• In local signaling, animal cells may communicate
by direct contact, or cell-cell recognition
 Plasma membranes

Gap junctions Plasmodesmata

between animal cells between plant cells

(a) Cell junctions

(b) Cell-cell recognition

• In many other cases, animal cells communicate
using local regulators, messenger molecules that
travel only short distances
• In long-distance signaling, plants and animals use
chemicals called hormones
Local and long-distance cell communication in animals

Local signaling Long-distance signaling

Target cell Electrical signal Endocrine cell Blood

along nerve cell vessel
triggers release of
neurotransmitter

Neurotransmitter
Secreting Secretory diffuses across
cell vesicle synapse Hormone travels
in bloodstream
to target cells

Local regulator
diffuses through Target cell Target
extracellular fluid is stimulated cell

(a) Paracrine signaling (b) Synaptic signaling

(c) Hormonal signaling

Structure of representative neurotransmitters
 Signaling Molecules and Their Receptors

• neurotransmitters
• Because neurotransmitters are hydrophilic,
– can’t cross the plasma membrane of target cells
– must bind to cell surface receptors.

• Many neurotransmitter receptors are ligand-gated

ion channels.
• Neurotranmitter binding opens the channels.
 Signaling Molecules and Their Receptors

• Abnormal autocrine signaling often contributes to

cancer.
• A cancer cell produces a growth factor to which it
also responds, thereby continuously driving its own
unregulated proliferation.
 The Three Stages of Cell Signaling: A Preview

• Cells receiving signals went through three processes:

– Reception
– Transduction
– Response

Animation: Overview of Cell Signaling, Cooper 15.1

Fig. 11-6-3

Overview of cell signaling

EXTRACELLULAR CYTOPLASM
FLUID
Plasma membrane

1 Reception 2 Transduction 3 Response

Receptor
Activation
of cellular
response
Relay molecules in a signal transduction pathway

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Signaling
molecule niso at a #

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 Reception:

A signal molecule binds to a receptor protein,

causing it to change shape

• The binding between a signal molecule (ligand)

and receptor is highly specific
• A shape change in a receptor is often the initial
transduction of the signal
• Most signal receptors are plasma membrane proteins
 Receptors in the Plasma Membrane

• Most water-soluble signal molecules bind to specific

sites on receptor proteins in the plasma membrane
• There are three main types of membrane receptors:
1. G protein-coupled receptors
2. Receptor tyrosine kinases
3. Ion channel receptors
 1- G protein-coupled receptors

• A G protein-coupled receptor is a plasma membrane

receptor that works with the help of a G protein

• The G protein acts as an on/off switch:

• If GDP is bound to the G protein, the G protein is
inactive
Hormonal activation of adenylyl cyclase
 Signaling-molecule binding site

1 2 3 4 5 6 7

Segment that
interacts with
G proteins

G protein-coupled receptor
 Membrane receptors—G protein-coupled receptors

G protein-coupled Plasma Inactive

membrane Activated Signaling molecule enzyme
receptor
receptor

GDP
G protein Enzyme GDP GTP
CYTOPLASM (inactive)

1 2

Activated
enzyme

GTP
GDP
Pi

Cellular response

3 4
 2. Receptor tyrosine kinases

• Receptor tyrosine kinases are membrane

receptors that attach phosphates to tyrosines
• A receptor tyrosine kinase can trigger multiple
signal transduction pathways at once
• Only three amino acids are phosphorylated
• Thyrosine
• Serine
• Threonin
 2. Membrane receptors—receptor tyrosine kinases
Signaling Ligand-binding site
molecule (ligand)
Signaling
molecule
 Helix

Tyr Tyr Tyr Tyr Tyr

Tyr
Tyrosines Tyr Tyr
Tyr Tyr Tyr Tyr
Tyr Tyr Tyr Tyr Tyr
Tyr

Receptor tyrosine
kinase proteins Dimer
CYTOPLASM

1 2

Activated relay
proteins

Cellular
Tyr Tyr P Tyr Tyr P Tyr Tyr P response 1
P
Tyr Tyr P Tyr Tyr P P Tyr Tyr P
Tyr Tyr P Tyr Tyr P P Tyr Tyr P Cellular
6 ATP 6 ADP response 2

Activated tyrosine Fully activated receptor

kinase regions tyrosine kinase
Inactive
relay proteins

3 4
 3. Ligand-gated ion channel

• A ligand-gated ion channel receptor acts as a

gate when the receptor changes shape
• When a signal molecule binds as a ligand to the
receptor, the gate allows specific ions, such as Na +
or Ca2+, through a channel in the receptor
3. Membrane receptors—ion channel receptors
1 Signaling
Gate
molecule closed Ions
(ligand)

Plasma
Ligand-gated membrane
ion channel receptor
2 Gate open

Cellular
response

3 Gate closed
 INTRACELLULAR RECEPTORS
• Some receptor proteins are intracellular, found in
the cytosol or nucleus of target cells
• Small or hydrophobic chemical messengers can
readily cross the membrane and activate receptors
• Examples of hydrophobic messengers are
– the steroid and
– thyroid hormones of animals
• An activated hormone-receptor complex can act as
a transcription factor, turning on specific genes
Signaling Molecules and Their Receptors

• Receptors for these molecules are members of the

nuclear receptor superfamily.
• They are transcription factors that have domains for
ligand binding, DNA binding, and transcriptional
activation.
• Proteins bind to certain regions of the DNA
– Helix turn helix,
– Helix Loop Helix
– Zip finger
– Leucine like region
• Ligand binding regulates their function as activators or
repressors of genes.
Signaling Molecules and Their Receptors

• Ligand binding has different effects on different

receptors.
• Some nuclear receptors are inactive in the
absence of hormone:
• Glucocorticoid receptor is bound to Hsp90
chaperones in the absence of hormone.
• Glucocorticoid binding displaces Hsp90 and
leads to binding of regulatory DNA sequences.
 Signaling Molecules and Their Receptors
• Peptide signaling molecules include peptide hormones,
neuropeptides, and polypeptide growth factors.

• Peptide hormones include insulin, glucagon, and

pituitary gland hormones (growth hormone, follicle-
stimulating hormone, prolactin, etc.).
Figure 15.3 Glucocorticoid action
 Signaling Molecules and Their Receptors
• Hormone binding may alter the activity of the receptor:

• In the absence of hormone, thyroid hormone receptor

is associated with a corepressor complex and
represses transcription of target genes.
• Hormone binding results in activation of transcription.
 Fig. 11-8-5
Hormone EXTRACELLULAR
(testosterone) FLUID

Plasma
membrane
Receptor
protein
Hormone-
receptor
complex

DNA

mRNA

NUCLEUS New protein

ANIMATION: Hormones
and Target Cell Receptors CYTOPLASM
Ch 31 in Biol
Figure 15.2 Structure of steroid hormones, thyroid
hormone, vitamin D3, and retinoic acid
Transduction:
Cascades of molecular interactions relay signals from
receptors to target molecules in the cell
• Signal transduction usually involves multiple steps
• Multistep pathways can amplify a signal: A few
molecules can produce a large cellular response
• Multistep pathways provide more opportunities for
coordination and regulation of the cellular response
 Signal Transduction Pathways

• The molecules that relay a signal from receptor

to response are mostly proteins
• Like falling dominoes, the receptor activates another
protein, which activates another, and so on, until the
protein producing the response is activated

• At each step, the signal is transduced into a

different form, usually a shape change in a protein

Animation: Signaling amplification, Cooper 15.3

 Protein Phosphorylation and Dephosphorylation

• In many pathways, the signal is transmitted by a

cascade of protein phosphorylations
• Protein kinases transfer phosphates from ATP to
protein, a process called phosphorylation
• Protein phosphatases remove the phosphates from
proteins, a process called dephosphorylation

• This phosphorylation and dephosphorylation

system acts as a molecular switch, turning
activities on and off
 A phosphorylation cascade
Signaling molecule

Receptor
Activated relay
molecule

Inactive
protein kinase
1 Active
protein
kinase
1
Inactive
protein kinase ATP
ADP Active P
2
protein
PP kinase
Pi 2
Inactive
protein kinase ATP
ADP Active P
3
protein
PP kinase
Pi 3
Inactive
protein ATP
ADP P
Active Cellular
protein response
PP
Pi
 Small Molecules and Ions as Second Messengers

• The extracellular signal molecule that binds to the

receptor is a pathway’s “first messenger”
• Second messengers are small, nonprotein, water-soluble
molecules or ions that spread throughout a cell by diffusion
• Second messengers participate in pathways initiated by
G protein-coupled receptors and receptor tyrosine kinases
• Cyclic AMP and calcium ions are common second messengers
Cyclic AMP

• Cyclic AMP (cAMP) is one of the most widely

used second messengers
• Adenylyl cyclase, an enzyme in the plasma
membrane, converts ATP to cAMP in response
to an extracellular signal

Adenylyl cyclase Phosphodiesterase

Pyrophosphate
P Pi

ATP cAMP AMP

• Many signal molecules trigger formation of cAMP
• Other components of cAMP pathways are G proteins,
G protein-coupled receptors, and protein kinases
• cAMP usually activates protein kinase A, which
phosphorylates various other proteins
• Further regulation of cell metabolism is provided by
G-protein systems that inhibit adenylyl cyclase
cAMP as second messenger in a G-protein-signaling pathway
First messenger

Adenylyl
G protein cyclase

G protein-coupled GTP
receptor
ATP
Second
cAMP messenger

Protein
kinase A

Cellular responses
 Pathways of Intracellular Signal Transduction

• In glycogen metabolism, protein kinase A

phosphorylates two enzymes:
• Phosphorylase kinase is activated, and in turn
activates glycogen phosphorylase.
• Glycogen synthase is inactivated.
• So, glycogen breakdown is stimulated and
glycogen synthesis is blocked.
Figure 15.21 Regulation of glycogen metabolism
by protein kinase A
 Pathways of Intracellular Signal Transduction

• Increased cAMP can activate transcription of

genes that contain a regulatory sequence—the
cAMP response element, or CRE.
• The free catalytic subunit of protein kinase A goes
to the nucleus and phosphorylates the transcription
factor CREB (CRE-binding protein).
• This leads to expression of cAMP-inducible genes.
• Protein phosphorylation is rapidly reversed by the action
of protein phosphatases, which terminates responses
initiated by receptor activation of protein kinases.
Figure 15.22 Cyclic AMP-inducible gene
expression
Figure 15.23 Regulation of protein
phosphorylation by protein kinase A and
protein phosphatase 1
 Pathways of Intracellular Signal Transduction

• cAMP can also directly regulate ion channels:

– It is a second messenger in sensing smells—
odorant receptors are G protein-coupled; they
stimulate adenylyl cyclase, leading to an increase
in cAMP.
– cAMP opens Na+ channels in the plasma
membrane, leading to initiation of a nerve impulse.
 Functions of Cell Surface Receptors
• G proteins have three subunits designated α, β, and γ.
• The α subunit binds guanine nucleotides, which
regulate G protein activity.
• In the inactive state, α is bound to GDP in a complex
with β, and γ.
• Hormone binding to the receptor causes exchange of
GTP for GDP. The α and βγ complex then dissociate
from the receptor and interact with their targets.
Figure 15.13 Regulation of G proteins
 Functions of Cell Surface Receptors
• A large array of G proteins connect receptors to
distinct targets.
• In addition to enzyme regulation, G proteins can
also regulate ion channels.
• Heart muscle cells have a different acetylcholine
receptor than nerve and skeletal muscle cells, which
is G protein-coupled.
• The α subunit of this G protein (Gi) inhibits
adenylyl cyclase. The Gi βγ subunits open K+
channels in the plasma membrane, which slows
heart muscle contraction.
 Calcium Ions and Inositol Triphosphate (IP3)

• Calcium ions (Ca2+) act as a second messenger in

many pathways
• Calcium is an important second messenger
because cells can regulate its concentration
The maintenance of calcium ion concentrations in an animal cell

EXTRACELLULAR
Plasma
FLUID
membrane

Ca2+ pump
ATP
Mitochondrion

Nucleus

CYTOSOL

Ca2+
pump
Endoplasmic Key
reticulum (ER)
Ca2+ High [Ca2+]
ATP pump
Low [Ca2+]
• A signal relayed by a signal transduction pathway
may trigger an increase in calcium in the cytosol
• Pathways leading to the release of calcium involve
inositol triphosphate (IP3) and diacylglycerol (DAG)
as additional second messengers

Animation: Signal Transduction Pathways

Pathways of Intracellular Calcium Signal Transduction
• Two major pathways of intracellular signaling use
second messengers derived from the membrane
phospholipid phosphatidylinositol 4,5-bisphosphate
(PIP2).
• Hydrolysis of PIP2 by phospholipase C produces two
second messengers: diacylglycerol (DAG) and inositol
1,4,5-trisphosphate (IP3).

• DAG remains associated with the plasma

membrane and activates protein-serine/threonine
kinases of the protein kinase C family.
• IP3 is a small polar molecule that is released to the
cytosol, where it signals release of Ca2+ from the ER.
Activation of phospholipase C by protein-tyrosine kinases
Ca2+ mobilization by IP3
Fig. 11-13-3

EXTRA-
CELLULAR Signaling molecule
FLUID (first messenger)

G protein

DAG

GTP
G protein-coupled PIP2
receptor Phospholipase C
IP3
(second messenger)

IP3-gated
calcium channel

Endoplasmic Various
reticulum (ER) Cellular
Ca2+ proteins
activated responses

Ca2+
(second messenger)
CYTOSOL
 The Effects of Sex Steroid Hormones
on CD38/ cADPR Signaling in Myometrium

Soner Dogan
University of Minnesota
H
Gq PLC
DAG
PIP2
IP3

VOC 2+
CaM
Ca
RyR IP3R
CaCaM
Ca2+
MLCK Myosin
Actin
Actomyosin
ATP
Contraction
Response: Cell signaling leads to regulation of
transcription or cytoplasmic activities

• The cell’s response to an extracellular signal is

sometimes called the “output response”
Nuclear and Cytoplasmic Responses

• Ultimately, a signal transduction pathway leads

to regulation of one or more cellular activities
• The response may occur in the cytoplasm or
may involve action in the nucleus
• Many signaling pathways regulate the
synthesis of enzymes or other proteins, usually
by turning genes on or off in the nucleus
• The final activated molecule may function as a
transcription factor
Fig. 11-14
Growth factor
Reception
Receptor

Phosphorylation
cascade
Transduction

CYTOPLASM

Inactive Active
transcription transcription
factor factor
Response
P

DNA

Gene

NUCLEUS mRNA
• Other pathways regulate the activity of enzymes
Cytoplasmic response to a signal: the stimulation of glycogen breakdown
by epinephrine
Reception
Binding of epinephrine to G protein-coupled receptor (1 molecule)

Transduction
Inactive G protein
Active G protein (102 molecules)

Inactive adenylyl cyclase

Active adenylyl cyclase (102)

ATP
Cyclic AMP (104)

Inactive protein kinase A

Active protein kinase A (104)

Inactive phosphorylase kinase

Active phosphorylase kinase (105)

Inactive glycogen phosphorylase

Active glycogen phosphorylase (106)

Response
Glycogen
Glucose-1-phosphate
(108 molecules)
 Fine-Tuning of the Response

• Multistep pathways have two important benefits:

– Amplifying the signal (and thus the response)
– Contributing to the specificity of the response

Signal Amplification
• Enzyme cascades amplify the cell’s response
• At each step, the number of activated products is
much greater than in the preceding step
The Specificity of Cell Signaling and Coordination of the Response

• Different kinds of cells have different collections of proteins

• These different proteins allow cells to detect and

respond to different signals
• Even the same signal can have different effects
in cells with different proteins and pathways
• Pathway branching and “cross-talk” further help
the cell coordinate incoming signals
 Fig. 11-17
Signaling
m olecule

Receptor

The specificity of cell signaling Relay

m olecules

Response 1 Response 2 Response 3

Cell A. Pathw ay leads Cell B. Pathw ay branches,

to a single response. leading to tw o responses.

Activation
or inhibition

Response 4 Response 5

Cell C. Cross-talk occurs Cell D. Different receptor

betw een tw o pathways. leads to a different response.
 Signaling Efficiency: Scaffolding Proteins and
Signaling Complexes
• Scaffolding proteins are large relay proteins to
which other relay proteins are attached
• Scaffolding proteins can increase the signal
transduction efficiency by grouping together
different proteins involved in the same pathway
Fig. 11-18

A scaffolding protein

Signaling Plasma
molecule membrane

Receptor

Three
different
protein
kinases
Scaffolding
protein
Termination of the Signal

• Inactivation mechanisms are an essential

aspect of cell signaling
• When signal molecules leave the receptor, the
receptor reverts to its inactive state
 Apoptosis (programmed cell death) integrates
multiple cell-signaling pathways
• Apoptosis is programmed or controlled cell suicide

• A cell is chopped and packaged into vesicles that

are digested by scavenger cells
• Apoptosis prevents enzymes from leaking out of a
dying cell and damaging neighboring cells
 Programmed Cell Death
Necrosis: Accidental cell death from acute injury.
Examples:
Hypoxia, temp change, toxins, physical and chemical factors.

Apoptosis: Programmed cell death; an active process.

old cells, lack of GFs, DNA damage, unwanted cells,
Protein misfolding in the endoplasmic reticulum

Characterized by:
– DNA fragmentation
– Chromatin condensation
– Fragmentation of the nucleus and cell
– No inflammation
 Apoptosis vs Necrosis
Apoptosis Necrosis
• Cellular fragmentation • No cell fragmentation
• No swelling • Cellular swelling (swell and lyse)
• Membranes remain intact • Membranes are broken
• Requires ATP • ATP is depleted
• Ladder-like DNA • DNA fragmentation is
fragmentation
random, or smeared
• In vivo, individual cells
appear affected • In vivo, whole areas of the
tissue are affected
• Cell is phagocytosed, no
tissue reaction • Inflammatory reaction
(No inflammation)
 Figure 17.1 Apoptosis

(C)
DNA from apoptotic cells.
 Programmed Cell Death

• Apoptotic cells and cell fragments are recognized and

phagocytosed by macrophages and neighboring cells,
and are rapidly removed from tissues.

• Necrotic cells swell and lyse; the contents are

released into the extracellular space and cause
inflammation.
• Apoptotic cells express “eat me” signals, such as
phosphatidylserine.
• In normal cells, phosphatidylserine is restricted to the
inner leaflet of the plasma membrane.
 Figure 17.2 Phagocytosis of apoptotic cells

In normal cells,
phosphatidylserine
is restricted to the
inner leaflet of the
plasma membrane.
 Programmed Cell Death

• Caspases are the ultimate executioners of

programmed cell death.
• They bring about the events of apoptosis by
cleaving 100 different cell target proteins.
• The activation of an initiator caspase starts a chain
reaction of caspase activation leading to death of
the cell.
 Targets of Caspases

Caspases cleave 100 different cell target proteins in

the event of apoptosis.
 Programmed Cell Death

• Ced-4 and its mammalian homolog (Apaf-1) bind

to caspases and promote their activation.
• In mammalian cells, caspase-9 is activated by binding to
Apaf-1 in a protein complex called the apoptosome.

• Cytochrome c is also required, which is released

from mitochondria.
Figure 17.5 Caspase activation
 Programmed Cell Death
• ced-9 in C. elegans is closely related to a mammalian
gene called bcl-2, which was first identified as an
oncogene.
• Bcl-2 inhibits apoptosis.
• Cancer cells are unable to undergo apoptosis.
• Mammalian cells encode about 20 proteins related to
Bcl-2, in three functional groups.
• Some inhibit apoptosis, while others induce caspase
activation.
• The fate of the cell is determined by the balance of activity
of proapoptotic and antiapoptotic Bcl-2 family members.
 Programmed Cell Death

• In mammalian cells, members of the Bcl-2 family act

at the mitochondria, which play a central role in
controlling programmed cell death.
• Cytochrome c is released from mitochondria, which
triggers caspase activation in the apoptosome.
Figure 17.8 The mitochondrial pathway of apoptosis
 Programmed Cell Death

• Caspases are also regulated by a family of

proteins called the IAP (inhibitor of apoptosis).
• They either inhibit caspase activity or target caspases
for ubiquitination and degradation in the proteasome.
• Regulation of programmed cell death is mediated by
signaling pathways, some acting to induce cell death
and others acting to promote cell survival.
• Many forms of cell stress, such as DNA damage, can
trigger programmed cell death.
Apoptosis of human white blood cells

2 µm
• Apoptosis evolved early in animal evolution and is essential
for the development and maintenance of all animals

• Apoptosis may be involved in some diseases (for

example, Parkinson’s and Alzheimer’s); interference
with apoptosis may contribute to some cancers
Effect of apoptosis during paw development in the mouse

Interdigital tissue 1 mm
Fig. 11-UN2
 You should now be able to:

1. Describe the nature of a ligand-receptor interaction

and state how such interactions initiate a signal-
transduction system
2. Compare and contrast G protein-coupled receptors,
tyrosine kinase receptors, and ligand-gated ion channels
3. List two advantages of a multistep pathway in the
transduction stage of cell signaling
4. Explain how an original signal molecule can produce a
cellular response when it may not even enter the target cell
5. Define the term second messenger; briefly describe
the role of these molecules in signaling pathways
6. Explain why different types of cells may respond
differently to the same signal molecule

7. Describe the role of apoptosis in normal development

and degenerative disease in vertebrates