The Progress of Cell Signaling

Schedule
Week Date Content

20230904 Cell communication and Signaling

2
20230906 GPCR-overview
20230911 GPCR-signaling
3
20230913 G protein and small G protein
20230918 RTK
4
20230920 GPCR and RTK crosstalk
20230925 Cytokine receptor
5
20230927 Ion channel
20231002 Nuclear receptor
6
20231004 MAPK pathway
20231009 PI3K and mTOR pathway
7
20231011 Protein modification
20231016 Ca2+ signal
8
20231018 cAMP signal
20231023
9 Drug development topics
20231025
 Reference Books and Link
• Biochemistry of Signal Transduction and Regulation. 3d ed. Gerhard Krauss,
2003
• Molecular Biology of the Cell, Bruce Alberts et.al., by Garland Science, Fifth
Edition 2008
• Molecular Cell Biology, Harvey Lodish et al，by W. H. Freeman and
Company，Fifth Edition 2003

• Biochemistrry, Lehninger
• The cell, Albert
• Cell, Lewis
• 细胞信号转导，孙大业主编
 Course Requirements
• Attendance: 10%
• Oral presentation: 30%
• Closed-book exam: 60%

Type Single-choice True or False Definition Total

(15 items, 2 points each) (20 items, 1 point each) (10 items, 5 points each)
Point 30 20 50 100
Cell Communication and Cell Signaling
Cells and the body

Cells are the basic units of life. Together, they form tissues that
themselves form organs, and eventually entire organisms.
 How a cell singals?

Each cells is
programmed to
respond to specific
stimulations of
extracellular signals.

Different signaling
cause different cell
activity and decide the
cell fate.
 Cell communication

All forms of communication between cells requires two

essential components:
1. a signaling cell that produces an instructive signal,
2. a responding cell that receives, acts on, and is
changed by the instructive signal.
Four main forms of cell communication
2) Contact signaling by plasma
1) Gap junction
membrane bound molecules

3) Chemical signaling 4) Exosome. ..

1) Gap junction
Ø Cells must be in direct contact
Ø Directly cell content exchange in both side of small
molecules (<1.5 kDa, inorganic ion or water soluble
small molecules, e.g. Ca2+, cAMP)
Ø Fast and reversible
e.g. Protein channel connecting the cytoplasm of two adjacent cells
2) Contact signaling by plasma membrane
bound molecules
Ø Cell-cell recognition by
interaction with molecules on cell
surface
Ø Also called contact-dependent
signaling or juxtacrine interaction
Ø e.g. immune response, cell
apoptosis

(major histocompatibility complex)

3) Chemical signaling
l Most chemical molecules are hydrophilic and only
some of them are hydrophobic.
l They can be proteins, peptides, amino acids and
other products.
l It can be both local or distal response.

Different forms for chemical signaling

transmission:
Ø Autocrine
Ø Paracrine
Ø Endocrine
Ø Chemical synapse signaling
3) Chemical signaling
Ø Autocrine

Autocrine signaling is a form of cell signaling in which a

cell secretes a chemical messenger that binds to the
receptors on that same cell, leading to changes in the cell.
3) Chemical signaling
Ø Paracrine
local signaling
Tissue fluid

Secretory Adjacent
cell target cell

Messenger molecules
e.g. cytokines, growth factors, neurotransmitters
3) Chemical signaling
Ø Slow, long-distance, long-term
Ø Endocrine Ø Blood transport, broad distribution
Ø Low ligand concentration
Endocrine cell Blood
vessel
Ø Receptor high affinity
Hormonal signaling: e.g. Insulin,
Thyroxine, Adrenaline…

Hormone travels in
bloodstream to target cells

Target cell
3) Chemical signaling
Ø Chemical synapse signaling

Neurotransmitter
Ø Fast e.g. acetylcholine,
Ø Local ligand serotonin, glutamate,
dopamine, γ-aminobutyric
acid (GABA)…
 Receptor “A”

Receptor “B”

Postsynaptic activity

Receptor “A” activity

Receptor “B” activity

4) Exosome
Nano-sized biovesicles released into surrounding body fluids upon
fusion of multivesicular bodies and the plasma membrane

Protein,
lipid,
mRNA,
miRNA,
et.al inside Intercellular communication by exosomes plays
a critical role in the regulation of cellular and
physiological processes.
migrasome (迁移体)
When cell migrate, the long fibrillar structures leaved behind are
retraction fibers, where small vesicles grow up, which is call
migrasome (Prof. Yu Li, Tsinghua University).
The production of migrasomes is highly correlated with the
migration of cells.

Cell Research (2022) 0:1 – 4 Cell Research volume 25, 24–38 (2015)
Front. Cell Dev. Biol., 16 June 2020
 Cell communication
1) Gap junction

2) Contact signaling by plasma membrane bound

molecules
3) Chemical signaling
Ø Paracrine
Ø Endocrine
Ø Autocrine
Ø Chemical synapse signaling
4) Exosome, migrasome…
ü Signal transduction is the From the cell From another
mechanism by which a cell itself cell
receives, acts on, and alters its
behavior in response to
outside
extracellular signal.
ü Signal transduction inside
pathway is the intracellular (cytosol)
processes that allow that
signal to be transduced from
the outer cell membrane of the
responding cell to the nucleus Cell response
where it effects a new program
of gene expression and cell
behavior.
 Three stages of cell signaling

1) Reception: ligand molecules are recognized by

receptor protein bound within a cell membrane. (Initiation)
2) Transduction: the interaction of receptor and ligand
leads to receptor conformation change, results in receptor
interacting with other intra-cellular molecules.
(Amplification/modulation)
3) Response stage is usually a cellular activity, as enzyme
catalysis, or the rearrangement of cytoskeleton (movement),
or specific gene activity. (Excution)
Overview of signal transduction pathways
 From the From the cell From another
environment itself cell

Light , Temperature,
Pressure, Taste, Smell,
Virus, Drug… outside

inside
Signal transduction is (cytosol)
essential for cell
communication;
it is also important for
the sensory and response
to the environment.
Cell signaling

Ø Extracellular signal
Ø Receptor
Ø Second messenger
Ø Signaling modulation or
Molecular switch
Ø Examples of receptor-induced
signaling pathway
Ø Characters of signaling
transduction
Ø Extracellular signal
Or signaling molecules (Ligand)
1) Lipid-soluble or water-soluble hormones
2) Nitric oxide (NO) and carbon monoxide (CO) as
cellular messengers
3) Cell surface receptors Light , Temperature,
Pressure, Taste, Smell,
4) Others Virus, Drug…

**Slow effect - alters gene expression

Intracellular Receptors
– Steroid Hormones, Thyroid Hormones, Retinoids and Vitamin D
**Fast effect - alters protein function
Nitric Oxide----Directly activates Enzymes
ØLigands
- Agonist
- Antagonist
Agonists are drugs or naturally occurring substances that
activate physiologic receptors, whereas antagonists are
drugs that block those receptors.
- Allosteric modulator: increases or decreases the strength
of the agonist-induced receptor response in a binding site
different from agonist
positive allosteric modulator, negative allosteric
modulator
Ø Characters of ligand binding to receptor

ü Specificity
ü High affinity
ü Saturation
ü Reversible
ü Affinity is controlled by receptor modification
Ø Receptor
Also refered as first messenger

ü Cell membrane receptor

• Ion channels
• G protein-coupled receptors (GPCR)
• Enzyme-Linked receptors
• Integrin
• others
ü Intracellular receptor
• Cytoplasmic receptor
• Nuclear receptor
Ø Ion channel

Ion channels are pore-

forming membrane proteins
which permit ions to cross
membrane including
plasma membrane and
organelle membrane.
Ø Ion channel
Ø G protein-coupled receptors

Ø Seven-helix transmembrane;
Ø N-terminal in the extracellular side and C-terminal in cytosol
Ø C-terminal: Ser- and Thr-rich
Ø GPCR phosphorylation can induce the desensitization and
recruiment of β-arrestin
Ø GPCRs: the most complex family of receptors
>800 members
Ø Enzyme-Linked Receptors

Six subfamilies:
① Receptor Tyrosine Kinases
② Tyrosine Kinase Associated Receptors
③ Receptor-like Tyrosine Phosphatases
④ Receptor Serine/Threonine Kinases
⑤ Receptor Guanylyl Cyclases
⑥ Histidine-Kinase Associated Receptors
Ø Receptor tyrosine kinases (RTKs)
Receptor tyrosine kinases recognize soluble or membrane bound
peptide/protein hormones that act as growth factors.

ØLigands
e.g. Vascular Endothelial Growth Factor (VEGF) family;
Platelet-derived Growth Factor (PDGF) family;
Transforming Growth Factor-b (TGF-b) family;
Neurotrophins;
Epithelial growth factor (EGF);
Fibroblast growth factor (FGF) family;
Insulin
Ø General Structure and Activation of RTKs
• RTKs generally exist as monomers with poorly active kinases.
• Binding of two ligands to the extracellular domains of two RTKs
forms or stabilizes an activated dimeric receptor.
• One kinase phosphorylates the other on a tyrosine residue in the
activation loop—autophosphorlation or tranphosphorylation,
resulting in kinase activation and dowstream signaling.
Ø Nuclear Receptors

Nuclear receptors are a class of proteins found within cells that are responsible
for sensing steroid and thyroid hormones and certain other molecules. In
response, these receptors work with other proteins to regulate the expression of
specific genes, thereby controlling the development, homeostasis, and
metabolism of the organism.
 Cell signaling

Receptor, first messenger

Second messenger
Second messengers are intracellular
signaling molecules released by the
cell in response to exposure to extra
cellular signaling molecules-
the first messengers.

Second messengers trigger physiological changes at cellular level such

as proliferation, differentiation, migration, survival, apoptosis and
depolarization.
 Small molecules and ions as
second messengers
p The extracellular signal molecule that binds to the receptor
is a pathway’s “first messenger”.

p Second messengers are small, nonprotein, water-soluble

molecules or ions that spread throughout a cell by
diffusion.

p Cyclic AMP and calcium ions are common second

messengers.

p Second messengers participate in pathways initiated by G

protein-coupled receptors and receptor tyrosine kinases
Ø Properties of second messenger
The intracellular “second messengers” are characterized by a
series of properties that make them particularly suitable as
elements of signal transduction:
- Intracellular messenger substances can be formed and
degraded again in specific enzyme reactions. Via enzymatic
pathways, large amounts of messenger substances can be
rapidly created and inactivated again.
- Messenger substances such as Ca2+ may be stored in special
storage organelles, from which they can be rapidly released by a
signal.
- Messenger substances may be produced in a location-specific
manner, and they may also be removed or inactivated according
to their location. It is therefore possible for the cell to create
signals that are spatially and temporally limited.
Ø Second messenger

- Hydrophobic character:
diacylglycerol or
phosphatidylinositol
- hydrophilic, phosphates
cytosolic:
cAMP, cGMP,
inositol
phosphates,
Ca2+

Low-molecular-weight messenger substances; Diffusible signal molecules

Cell signaling

Receptor, first messenger

Second messenger

Kinases

Signaling modulation or
Molecular switch
Ø Protein phosphorylation
and dephosphorylation
Ø GTPase switch proteins
Ø Signaling modulation or Molecular switch
Protein phosphorylation and dephosphorylation
ü Protein kinases transfer
Signaling
phosphates from ATP to molecule Receptor

protein, a process called Activated relay molecule

Inactive
phosphorylation. protein kinase
Active
1
protein kinase
ü Phosphorylation can flip 1
Inactive
protein kinase ATP
a protein from “active” 2
ADP Active
P
protein kinase
PP
Pi 2
to “inactive” or vis-versa. Inactive
ATP
protein kinase ADP Active P
ü In many pathways, the 3
PP
protein kinase
Pi 3

signal is transmitted by a Inactive

protein ATP
P
ADP Active Cellular
cascade of protein Pi
PP
protein response

phosphorylations.
Ø Signaling modulation or Molecular switch
guanosine triphosphatases (GTPase, 鸟苷酸三磷酸酶)
Guaninenucleotide exchange factor,
GEF, 鸟苷酸交换因子

G protein acts as a molecular switch

during signal transduction. After
binding to GDP, the G protein is in
an inactive state. After GTP replaces
GDP, G protein activates and
transmits signals
GTPase activating protein, GAP,
鸟苷酸三磷酸酶激活蛋白
Ø Examples of receptor-induced signaling pathway

l GPCR signaling
- cAMP pathway
- Phosphatidylinositol pathway
l RTK signaling
- MAPK pathway
- PI3K/Akt pathway
Ø GPCR-coupled adenylyl cyclase-cAMP system

- Gs-coupled GPCRs,
cAMP↑
- Gi/o-coupled GPCRs,
cAMP↓
Ø GPCR Phosphatidylinositol (磷脂酰肌醇) Pathway

- Gq-coupled GPCRs

Ø IP3/Ca2+ and DAG/PKC pathway

Ø The Effectors of GPCRs

β-arrestin
Ø RTK signaling

MAPK (Mitogen-activated
Protein Kinase) Cascades

PI3K/Akt
pathway
 Ø RTK signaling
- MAP kinase pathways
Activated Ras induces a kinase
signal cascade that
culminates in activation of
MAP kinase.
MAP kinase is a
serine/threonine kinase that
can translocate into the
nucleus and phosphorylation
of many different proteins,
including transcription
factors that regulate gene
expression.
Ø RTK signaling
- PI3K/Akt pathways
 Insulin receptor induced PI3K Signaling

IRS: insulin receptor substrate

Berridge MJ. Cell Signalling Biology. 2014. doi:10.1042/csb0001002

Ø Insulin and glucagon work together to maintain a
stable blood glucose level

胰高血糖素
Summery of four classes of receptors and signaling
Ø Character of signaling transduction
ü Convergence; Divergence;

Converge on Ras Divergence

Ø Character of signaling transduction
ü Cascade response; Crosstalk

GPCR and RTK

Cascade response signaling Crosstalk
Ø Character of signaling transduction
Ø Conserved regulation:
- e.g. similar regulation through phosphorylation and
dephosphrylation in Ser, Thr and Tyr; conserved in evolution
among species
IR

PI3K
PTEN

FOXO

FOXO
apoptosis

Humans C. elegans
Summary
Cell signaling
Ø Extracellular signal (Ligand)
Ø Receptor (membrane and intracellular receptor)
Ø Second messenger (e.g. cAMP, IP3, Ca2+….)
Ø Signaling modulation or Molecular switch
(Phosphorylation, GTPase)
Ø Examples of receptor-induced signaling pathway
(GPCR: cAMP; IP3-Ca2+; DAG-PKC
RTK: MAPK, PI3K/Akt pathway)
Ø Characters of signaling transduction
(Convergence; Divergence; Cascade response;
Crosstalk; Conserved regulation)
 Evolution and properties of cell
signaling cascades

Past: Now:
Enumerate Modules
components Design Logic
Cross-talk

Specificity
Affinity

Cooperativity
Sensitization
Amplification
Integration

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
 Cell signaling in Eukaryotes
p Eukaryotic signaling systems are much more elaborate
than those in yeasts or bacteria (Prokaryotes).

p Flies, worms and mammals all use essentially similar

machinery for cell communication.

p In plants, as in animals, cells are in constant

communication with one another. Plants use different
signaling molecules and receptors than animals.

p More than 1500 genes encode different receptor

proteins in human.
 Overview: the cellular internet

p Cell signaling is essential for life of unicellular and

multicellular organisms.

p Cells can communicate to the cell(s) next to them or

cells from much further away in another part of the body.

p How do cells send and receive a signal?

p Once cells receive the signal, how does this lead to

changes inside the cell (response)?
 Significance of cell signaling
üFor normal functioning coordination of every signaling
pathway is necessary

ü Defect can be in any component of signaling ultimately

leading to the disease development.

ü C e l l s i g n a l i n g h a s b e e n i d e n t i f i e d i n C a n c e r,
Cardiovascular diseases (hypertension, heart disease, etc.…),
Alzheimer's disease, and many other disorders.

üCell Signaling – an important area of research for drug

discovery
 Future of cell signaling

ü With advances in separations methodology, mass spectrometry,

and hybridization, the complex protein interactions in cellular
signaling networks should become clear.

ü Future cell signaling studies would involve integration of

genomic, transcriptomic, proteomic, and metabolomic data that
will provide complete picture of cellular mechanisms and their
responses.
 Schedule
Week Date Content

20230904 Cell communication and Signaling

2
20230906 GPCR-overview
20230911 GPCR-signaling
3
20230913 G protein and small G protein
20230918 RTK
4
20230920 GPCR and RTK crosstalk
20230925 Cytokine receptor
5
20230927 Ion channel
20231002 Nuclear receptor
6
20231004 MAPK pathway
20231009 PI3K and mTOR pathway
7
20231011 Protein modification
20231016 Ca2+ signal
8
20231018 cAMP signal
20231023
9 Drug development topics
20231025
The end!