No cell is an island

z Cells communicate with each other

z Cells send and receive information (signals)
z Information is relayed within cell to produce a
Cell-to-Cell Signaling response (signal transduction)
Receptors

Signaling process Life is not easy

z Recognition of signal z The effects of activation of cell surface receptors
z Receptors are more complicated than a simple step-by-
z Transduction step cascade
z Change of external signal into intracellular message z Extensive networking and cross talk
z Effect z By no means signaling is a linear event
z Modification of cell behavior z Key is integration

Recognition Transduction

z Performed by receptors z Activated receptor interacts with other

z Ligand will produce intracellular signaling molecules and
z Directly changes cellular activity
response only in cells
or
that have receptors
z Produces some intermediates (second messengers)
for this particular that change cellular activity
ligand
z Each cell has a specific
set of receptors

1
Cellular effects
z Metabolism
z Gene expression
z Growth and differentiation (cell cycle)
z Secretion
z Structure (cytoskeleton)
Receptors
z Molecules specialized in recognizing signals
z Can be inside the cell or out
z Specific for a signal (they will recognize only a
signal that they are made for)
z Energy from the signal (stimulus) or ligand
binding alters the structure of the receptor
To be a receptor a molecule
z Has to have an endogenous ligand
z Has to bind it with high affinity (strength)
z Has to recognize the biologically active ligand
from other similar molecules (specificity)
z Has to produce the biological response
All the conditions have to be met!!!
Why so complicated?
z Amplification
z Reliablity
z Redundance
Receptors
z Ligand – substance that binds to receptor with
high affinity
z Binding of the ligand produces biological response
z Ligands can be endogenous or exogenous
z Agonist – biological response of the same type as
the endogenous ligand (can be smaller)
z Antagonist – binds and blocks the response
Definition of a receptor
z Molecules within or on the surface of target cells
to which a substance endogenous to the
organism attaches itself with high affinity and
specificity as an essential step in modifying
cellular function
2
Characterization of the receptors
based on localization Intracellular receptors

z Intracellular receptors z If a substance can cross the membrane it will

z For molecules that can bind to intracellular receptors
enter the cell z Localized in nucleus or cytoplasm
z Those in cytoplasm translocate to the nucleus upon
activation
z Plasma membrane z Ligand activated transcription factors
receptors z Genomic effects
z For molecules that can not

Domain structure of nuclear

Structure of nuclear receptors receptors

z Superfamily of ligand-activated transcription

factors
z Bind to specific DNA sequences as dimers
z Similar structure and high homology
z Two highly conserved regions

Highest homology region –

zinc finger motif

Model of activation of intracellular

Ligands of intracellular receptors receptors

z Small hydrophobic molecules that can cross the z In the absence of

membrane hormone, transcriptional
z Steroid hormones activation is inhibited
z Thyroid hormones z Hormone binding
z 1,25-dihydroxychole- relieves inhibition
calciferol (vitamin D3) z Release of inhibitory
z Retinoic acid complex – hsp
z Exposing of DNA
binding domain

3
Mechanism of intracellular (nuclear) Consensus sequences (Hormone
receptor action Response Elements)

z Ligand binding activates the

receptor Estrogen receptor
z Activated receptor binds to
regulatory DNA sequences
called Hormone Response
Elements Retinoic acid receptor
z Localized in the 5’ flanking
regions of target genes
z N-terminus has a transcription
activation domain

Why do we need cell surface

Structure of nuclear receptors receptors?

z Work as dimers z Most signals can not cross the membrane

z Directly activate z It can’t work if it can’t get in
transcription (ligand z But it can send a messenger
activated transcription z When activated, a receptor on the surface
factors) “passes” the signal to intracellular second
messengers

Second messengers Plasma membrane receptors

z Molecules inside a cell that z About 20 families so far

z Change of function of the cell z Based on similarities in structure, ligand binding
z Signal to the nucleus for genomic effects and signal transduction strategies

4
Plasma membrane receptors
z Different sizes and shapes
z Different isoforms within the family arose by
gene duplication and divergence
z Selective expression decides about physiology
of the cell
z Ligand binding or physical energy (light,
pressure, temperature, voltage) activate the
receptor
Signaling strategies
z Receptors that have intrinsic
enzymatic activity (they are enzymes)
z Receptor tyrosine kinases
z Receptors that activate intracellular
second messengers
z G protein coupled receptors
z Cell adhesion receptors
Ligands that use ion channel
receptors
z Histamine
z Serotonin
z Heat and cold
z Mechanical deformation of membrane
Signaling strategies
z Ion channels receptors
z Ligand gated ion channels
z Voltage gated ion channels
z Receptors that are linked to
cytoplasmic enzymes
z Cytokine receptors
Ion channels receptors
z Receptor molecule is an ion channel at the
same time
z Ligand gated ion channels
z Voltage gated ion channels
z Activation of receptor opens a channel and
changes excitability of the membrane
Signaling strategies
z Ion channels receptors
z
z Ligand gated ion channels
z
z Voltage gated ion channels
z Receptors that are linked to
cytoplasmic enzymes
z Cytokine receptors
5
Cytokine receptors Cytokine receptors

z Receptors for interleukins and polypeptide z All have 4 subunits

growth factors
z Do not have enzymatic activity but bind
cytoplasmic protein tyrosine kinases
z Signal to nucleus
z Regulate gene expression
z Homodimer or
heterotrimer/heterodimer
z Single transmembrane region
z Extracellualr fibronectin
domain (ligand binding site)
z Cytosolic domain binds JAKs (kinases)

Ligands Cytokine receptors activation

z Interleukins z Dimerization or conformational change

z Growth hormone z That brings together two JAKs
z Prolactin z JAKs cross-phosphorylate
z Erythropoietin z Activated JAKs phosphorylate (activate) STATs
(transcription factors )
z STATs translocate to the nucleus

Signaling to the nucleus by

JAK/STAT pathway Signaling strategies

z Receptors that have intrinsic

enzymatic activity (they are enzymes)
z Guanylyl cyclase
z Receptor tyrosine kinases

z Receptors that activate intracellular

second messengers
z
z G protein coupled receptors

6
Receptor tyrosine kinases (RTK) Receptor tyrosine kinases (RTK)

z Cell-surface receptors with intrinsic enzymatic

activity (tyrosine kinase)
z Cytoplasmic domain has tyrosine kinase activity
z Mostly receptors for peptide/protein growth
factors
z Regulate growth and differentiation and cell
survival
z Single transmembrane domain
z Usually single polypeptide
z Insulin family is an exception
z Extracellular binding site has
immunoglobulin or fibronectin
domains
z Intracellular kinase domain includes
tyrosines that can cross-
phosphorylate

Subfamilies of receptor tyrosine Ligands of receptor tyrosine kinases

kinases (RTK) (RTK)

z EGF
z Insulin
z PDGF
z FGF
z NGF

Activation of RTKs Activation of RTKs

z Ligand binding promotes conformational change z Dimerization activates

and dimerization of the receptor enzymatic activity of the
z Some RTK are receptor molecule (tyrosine
dimers (insulin kinase)
receptor) but z Cross-phosphorylation of the
ligand binding is cytosolic domain on the
necessary for partner
activity

7
 Ligand binding leads to
Activation of RTKs
autophosphorylation of RTKs

z Phosphotyrosine residues interact with proteins

that have SH2 and PTB domains
z Two mechanisms of interactions with the effector
z Activation of enzymes with SH2 domains
z Through adapter protein

Guanylyl cyclase receptor Guanylyl cyclase receptor

z Another receptor with intrinsic enzymatic activity z Receptor catalyzes formation of cGMP from
(guanylyl cyclase) GTP
z Binding of the ligand activates the enzymatic z cGMP regulates
activity of the receptor molecule z cGMP gated ion
z Receptor for atrial naturetic factor and other channels
peptides z cGMP activated
protein kinases
z cGMP-phosphodiesterases

Signaling strategies Seven-helix receptors

z Receptors that have intrinsic z A.k.a. G protein coupled receptors

enzymatic activity (they are enzymes) z Plasma membrane receptors
z
z Guanylyl cyclase
z Involved in a range of signaling pathways,
z
z Receptor tyrosine kinases including light detection, odorant detection, and
detection of certain hormones and
z Receptors that activate intracellular neurotransmitters
second messengers
z G protein coupled receptors

8
Molecular properties of seven-helix Molecular properties of seven-helix
receptors receptors

z Seven transmembrane z C-terminus faces cytosol and

regions of 22-24 has phosphorylation sites
hydrophobic residues z Place for regulation
z N-terminus faces outside z A cytosolic loop between
z Often involved in activation helices 5 and 6 is the place
or ligand binding for interaction with G protein

Hormones that act through seven-

Ligand binding helix receptors

Different possibilities for ligand binding z Epinephrine and norepinephrine

1. Ligand binding site in a pocket among z Glucagon
extracellular ends of the helices (1/3 into the z Bradykinin
membrane) z ACTH
2. N-terminal binding z Vasopressin
3. Truncated N-terminus activates the receptor z Luteinizing hormone LH
z Thyroid stimulating hormone TSH

Seven-helix receptors Signaling by seven-helix receptors

z Activated receptor catalyzes G protein z A typical signal transduction cascade comprises

activation z G protein
z Exchange of nucleotide bound to G protein z Enzyme
(replacement of GDP with GTP) z Second messengers
z Activated G protein initiates signaling cascade z Target protein - effector
z Not necessary all steps are involved!!!!

9
Other receptors Cell adhesion receptors

z Receptor serine/threonine kinases

z TNF receptor family
z Notch receptors
z Hedgehog receptors

To be discussed later

10