Chapter 16

Cell Communication

Extracellular signal molecules can act over

short or long distances

Figure 16-3

Overview of signal initiation and responses

1.

Receptor recognizes
stimulus (3 types
shown)

2.

Signal transferred to
cytoplasmic surface of
receptor

3.

Signal transmitted to
effector molecule

4.

Cessation of response

Signal transduction - signal conversion

First messenger

Effector
Second messenger

Figure 16-13

Same signal molecule can induce different

responses in different target cells

Although heart and salivary gland have same

receptor, different intracellular effector proteins
are activated
Figure 16-5

Extracellular signals bind to cell-surface or intracellular

receptors

Carrier protein

Figure 16-8

Animal cells get multiple signals

Figure 16-6

Extracellular signals can act rapidly or slowly

Fig. 16-7

Signaling proteins can relay, amplify, integrate

and distribute incoming signal
First messenger

Second messenger

signal transduction -- stimulus

received by cell-surface
receptor is different than
signal released in cell interior
signal pathways -- series of
distinct proteins that alter the
conformation of the
downstream protein

Figure 16-13

Second messengers

small, nonprotein intermediaries acting in signal

transduction
e.g., cAMP, calcium, lipid-derived

3 classes of cell surface receptors

1. Ion channel coupled receptors

Figure 16-17a

3 classes of cell surface receptors

2. G-protein-coupled receptors

Figure 16-17b

3 classes of cell surface receptors

3. Enzyme-coupled receptors

Figure 16-17c

Basic characteristics of
cellsignaling systems

Many intracellular signaling molecules act

as molecular switches

shape change by phosphorylation

Figure 16-15

Protein phosphorylation
may cause a conformational
change in the protein
may be part of a protein
binding site
may be added to serine,
tyrosine, or threonine
may inc. or dec. proteins
activity

most substrates are other

enzymes
Protein kinases/phosphatases
change the shape/activities of
the proteins they modify
continual balance of kinase
and phosphatase activities
human genome ~500
different kinases and ~100
different phosphatases

Many intracellular signaling molecules act

as molecular switches

shape change by GDP/GTP exchange

G-proteins are NOT kinases
Figure 16-15

The activity of monomeric GTP-binding proteins is

controlled by two types of regulatory proteins

G-proteins are
NOT kinases

GAP - GTPase-activating proteins

GEF - Guanine nucleotide-exchange factors

Figure 16-16

2 alternate types of
signal transduction
pathways
1. G protein-linked
receptor
2. Protein kinase receptor

All GPCRs have a similar structure

7 transmembrane -helices

Figure 16-18

G protein-coupled receptors (GPCRs)

e.g., adenylyl cyclase

e.g., glucagon receptor

e.g., cAMP

GPCR are largest superfamily of proteins in animal genome

(e.g., nematode has 19,000 genes of which 1000 are GPCRs)
Target of ~40% of modern medicinal drugs
See Figure 16-19

G-protein activation

Heterotrimeric G proteins

1.

signal causes shape change

in receptor

2. G protein binds to receptor

3. GTP binds to G protein,
causing shape change
4.

subunit and subunits

separate

Fig. 16-19

Signal relay to effector

activated subunit
changes effector
shape (on)

Fig. 16-20

Ending the response

Turning off the
effector
1.

GTP hydrolyzed and

subunit changes shape
(off)

2. G protein no longer
binds effector, but
reforms trimer
3. Effector changes
shape (off)

Fig. 16-20

Ending the response - II

1.

Receptor
desensitization

GRK (G protein-coupled receptor kinase)

phosphorylates receptor

2. arrestin binds, blocks G protein binding

3. receptor endocytosed from PM

G-protein signaling

--Shows that response can be ended by GTP hydrolysis

and endocytosis of receptor

Specificity of G protein-coupled responses

not all parts of signal transduction machinery identical in
all cells
multiple forms of receptors (e.g., 9 different isoforms
of epinephrine receptors) with different ligand and G
protein affinities
multiple G proteins ( 20 G; 5 G; 11 G ); various
combinations
Gs stimulatory and Gi inhibitory

G proteincoupled receptors
--some G proteins directly regulate ion channels

Some G proteins directly regulate ion channels

K+

16_19_open_K_chan.jpg

subunits active
slows heart beat

See Fig. 16-21

G proteincoupled receptors
--some G proteins activate membranebound enzymes

Enzymes activated by G proteins catalyze production of

small intracellular signaling molecules
e.g. Adenyl cyclase and phospholipase C

e.g. cAMP; IP3/DAG

Figure 16-20

Glucose is stored in animal cells as

glycogen (polymer of glucose)
Glucagon & epinephrine

Insulin

(glycogen) phosphorylase makes more glucose

glycogen synthase makes more glycogen

Glucagon (pancreas)

glucagon and epinephrine blood glucose by

inhibiting glycogen synthase and activating
phosphorylase
the 2 hormones bind to different receptors, yet
lead to same intracellular response, How?? Both
use the same secondary messenger cAMP.

Adenylyl cyclase

integral membrane protein

that makes cAMP
an effector
See Fig. 16-23

Adrenaline stimulates glycogen breakdown in

skeletal muscle cells

Protein kinase A (PKA) is

activated by cAMP
PKA also phosphorylates
glycogen synthase, which
inhibits glycogen synthesis

Figure 16-25

Adrenaline in the same cells also stimulates glucose synthesis

cAMP can activate

gene transcription

Enzymes needed for

gluconeogenesis

Figure 16-26

cAMP Signaling

Signal amplification

Each GPCR activates

multiple G proteins
Each active adenylyl
cyclase make many cAMPs
Each active PKA can
phosphorylate multiple
phosphorylase kinases
Etc..

Reversal of signal
phosphatase-1 removes the
phosphates that were added by
PKA
cAMP is destroyed by cAMP
phosphodiesterase

Figure 16-23

Phosphatidylinositol (PI) and phosphoinositides (PIPs)

PI

PIPs
Numbering
the carbons in
inositol
Phosphorylation
of OH in inositol
Named according to
ring position (in
parentheses) and
number of phosphate
groups (in subscript)

Animal cells have several PI and PIP kinases

and phosphatases

PI headgroups are recognized by protein

domains that discriminate the different forms

Proteins are recruited to regions of the

membrane where these PIs are present

Enzymes activated by G proteins catalyze production of

small intracellular signaling molecules
e.g. Adenyl cyclase and phospholipase C

e.g. cAMP; IP3/DAG

Figure 16-20

Lipid-derived second messengers

inositol

phospholipases -hydrolytic enzymes that

split phospholipids
PLA -- phospholipase A
PLD -- phospholipase D
PLC -- phospholipase C

Phosphatidylinositol-derived
are best studied

Phosphatidylinositol (PI)- mediated responses

phospholipase C
cleaves PIP2 into DAG
and IP3

DAG

IP3

See Fig. 16-25

How does PLC get activated?

See Fig. 16-25

Diacylglycerol (DAG)
lipid molecule remains in PM
recruits and activates protein kinase C (PKC)
PKC is a serine/threonine kinase that is imp. in many
cellular events (e.g., cell growth, differentiation,
metabolism and transcriptional activation)
phorbol esters -- plant compounds that mimic DAG
and activate PKC; cells lose growth control and
behave as malignant cells

IP3 (inositol 1,4,5-trisphosphate)

small, water soluble
binds to a Ca++ channel receptor
on smooth endoplasmic reticulum
(SER); Ca++ release

Phospholipase C releases lipid-derived second messengers

Figure 16-27

Ca++ as an intracellular messenger

Ca++ release from intracellular stores acts as a second
messenger
First messenger: hormones, neurotransmitters, electrical
activation (muscle)

Calcium Signaling

Ca++ is not made enzymatically

[Ca++] controlled by pumps and channels
Free intracellular [Ca++] ~ 0.1 M
[Ca++] within lumen of ER ~ 1 mM (10,000-fold
higher)
channels into
cytoplasm normally
closed
out of cytoplasm
pumps usually active

Where are Ca++ release channels found?

IP3Rs: present on
smooth ER of most
cell types

Unlike cAMP, Ca++ modulates a broad range

of effectors
cAMP acts on kinases; primarily PKA
Ca++ can activate kinases, ion pumps,
proteases, phosphatases, Ca ++-binding
proteins, etc.
Calmodulin -- best known Ca ++-binding
protein

Calmodulin (CaM)
binds Ca++ only in
stimulated cells
(affinity too low)
Ca++ binding changes
confirmation and
affinity for other
proteins
Activated CaM
activates Ca++ pump
to reduce cytosolic
levels

Figure 16-29

Calmodulin

Role of GPCR in vision

cGMP keeps
open

Receptor - rhodopsin
G protein -transducin
Effector - cGMP
phosphodiesterase
response 20 msec
(20/1000th of a sec)
See Figure 16-30

16_29_amplifies_light.jpg

See Fig. 16-31

2 alternate types of
signal transduction
pathways
1. G protein-linked
receptor
2. Protein kinase receptor

Receptor tyrosine kinases (RTKs)

transmembrane receptors
enzymes that adds phosphate groups to
tyrosine aa on proteins
each RTK monomer crosses PM once (GPCR
7Xs)
> 50 different RTKs have been identified

Typical RTK

1.

Inactive receptor monomers

2. Ligand induces dimerization and

autophosphorylation
3. Signaling protein bind to specific
phosphorylated Tyr
Figure 16-32

Important concepts
phosphotyrosine motifs -- only
tyrosines surrounded by certain aa are
phosphorylated
SH2 domains (src homology 2) and
PTB domains (phosphotyrosine binding);
sites in proteins that recognize the
phosphorylated tyrosines; found in
many cell signaling proteins

Interaction between SH2 domain and

phosphotyrosine

~ 100 aa long
Found in ~110 different human proteins
SH2 compact plug-in module can be inserted nearly
anywhere in a protein sequence and not disturb protein
folding

Role of RTK in cellular activities

many types, but best studied:
-- hormones - e.g. insulin, growth hormone
-- growth factors - epidermal growth
factor (EGF), fibroblast growth factor
(FGF), platelet-derived growth factor
(PDGF)

Insulin receptor

tetramer of two and two polypeptide chains

insulin binding to changes conformation of
activated receptor phosphorylates self
(autophosphorylation) and soluble proteins (insulin
receptor substrates; IRSs)

PTB domain of IRS-1

binds to insulin receptor
receptor phosphorylates
docking sites on IRS-1

IRS-1 (insulin receptor substrate-1)

docking sites

pTyr binding on
insulin receptor

SH2 domain

RTK activate PI-3-kinase

Fig. 16-33

Phosphoinositides direct major signaling cascades

-can act as docking sites and as secondary messengers
PI

PIP

PIP2

PIP3 -- PI(3,4,5)

phosphorylated inositol rings bind to modules on many

proteins
recruits specific protein to cytoplasmic face of PM
PH: pleckstrin homology domains targets proteins to
membranes by binding PIP2 or PIP3 when they are
produced

PIP3 Produced in Response to Stimulation of a RTK

e.g., PDGF or Insulin

-GFP fused to a PH (PIP3 binding domain from a tyr kinase)

-PDGF causes the production of PIP3 at the plasma membrane

PH = pleckstrin homology
Dr. Seth Field

PI-3-kinase and PKB activation

Akt = Protein kinase B

Fig. 16-33

PI3K generates PIP2 and PIP3

protein kinase B (PKB)
binds/partially-activated by PIP2
and PIP3 via its PH domains
fully activated by another
kinase(s)
signal reversed by
dephosphorylating receptor
Inc. protein syn.
Inc. glucose uptake
Inc.

glycogen syn.

thus, insulin
leads to lower
blood glucose

PH domains -pleckstrin homology

domain -- binds
PIP2/PIP3

The importance of Ras

Ras is a monomeric G protein held to PM by
lipid group
Monomeric G proteins mimic G in function
nearly all RTK activate Ras
30% of human tumors have a mutated ras (or
ras-like) gene

Most RTK activate the monomeric G protein Ras

Grb2

Sos

Ras GEF

MAP kinase
pathway

Ras mutations leading to tumors

prevent GTP hydrolysis; Ras always
on
Figure 16-33

Ras

MAP kinase cascade

Mitogen-activated protein
kinase
growth factor binding leads to
inc transcription of genes
involved in growth responses
Ras GTP hydrolysis is induced
by a GTPase activating protein
(30 min dec to 1/10 sec)
Ras mutations leading to
tumors prevent GTP hydrolysis;
Ras always on

MAP kinase pathway

16_32_MAP-kinase.jpg

See Fig. 16-34

Signals that originate from ECM

integrins transmit signals that

influence cell growth, shape,
migration, differentiation and survival
normal cells do not grow in
suspension

Signals induce cell growth

1.

clustering of
integrins induces
protein binding

2. P-tyr activation of
src and FAK kinases
3. Ras activated
4. MAP kinase cascade
inc. transcription of
genes involved in
growth

FAK = Focal adhesion kinase

src is an oncogene

Actin
P-tyr proteins

Extracellular signals bind to cell-surface or intracellular

receptors

Carrier protein

Figure 16-8

Role of nitric oxide as an intracellular messenger

Mode of action for Viagra

smooth muscle cell

around arterial
blood vessels

NO (nitric oxide) released by nerve and endothelial cells in penis

NO --> higher cGMP levels --> muscle relaxation and increased
blood flow --> erection
Viagra inhibits PDE5 (form of cGMP phosphodiesterase; found only
in penis) keeping cGMP levels high

Animal cells get multiple signals

Figure 16-6

Signaling cascades are not just linear

convergence - 2 ligands, 1 path

divergence - 1 ligand, many paths
divergence

convergence

Figure 16-40

Scaffolds help organize cascades

structural not enzymatic components

provide spatial localization & substrate
specificity