SIGNALING

Extracellular/Intercellular Intracellular
Signaling by extracellular, secreted molecules Cell's direct signaling

• Autocrine • Juxtracrine
• Paracrine
• Endocrine
 Signaling Molecules (Signals/Ligands)
• The ligand is the primary messenger.
• As the result of binding the receptor, other molecules or second
messengers are produced within the target cell.
• Second messengers relay the signal from one location to another (such as
from plasma membrane to nucleus) leading to cascade of events/changes
within a cell
• Messenger molecules may be amino acids, peptides, proteins, fatty acids,
lipids, nucleosides or nucleotides.
• Hydrophilic messengers bind to cell membrane receptors.
• Hydrophobic messengers bind to intracellular receptors which regulate
expression of specific genes.
• A ligand binds its receptor through a number of specific weak non-covalent
bonds by fitting into a specific binding site or "pocket ".
• In situations where even low concentrations of a ligand will result in binding
of most of the cognate receptors, the receptor affinity is considered to be
high.
• Low receptor affinity occurs when a high concentration of the ligand is
required for most receptors to be occupied.
• SIGNAL With prolonged exposure to a ligand (and occupation of the receptor)
cells often become desensitized.
1) Hormone
• a class of signalling molecules produced
• by glands in multicellular organisms.
• transported by the circulatory system to target distant
organs to
• regulate physiology and behaviour.
2) Neurotransmitter
• endogenous chemicals that transmit signals across a
synapse
• from one neuron to another "target" neuron.
• released from synaptic vesicles from synapses into the
synaptic cleft, where received by receptors on other
synapses.
3) Cytokines
• signalling molecule of immune system.
 Steroid hormones and nuclear receptor superfamily

• Receptor- Intracellular proteins

(cytosol/nucleus)
• Able to diffuse plasma membrane
• Testosterone, vitamin D3, retinoic acid etc
• Bind intracellular receptors and regulate gene
expression
 Nitric oxide (NO) and Carbon Monoxide (CO)

• NO- major paracrine signaling molecule (short half

life)
• Acts in Nervous, immune and circulatory system
• Able to diffuse plasma membrane
• Alters the activity of intracellular enzymes
• Main target guanylyl cyclase – stimulate synthesis of
cGMP– role in blood vessel dilation
• Also done nitrosylation at cysteine residues in some
proteins
 Neurotransmitters
• Carry signals b/w neurons and neurons to
target cells
• E.g. acetylcholine, dopamine, epinephrine,
serotonin, GABA (gamma- aminobutyric acid)
• Hydrophillic - so do not cross membrane
• Paracrine signaling
• Bind to their respective receptors- ion
channels or GPCRs
Peptide hormones and Growth factors
 Eicosanoids
• Lipids signals
• E.g prostaglandins, prostacyclin,
thromboxanes, leukotrienes
• Autocrine or paracrine signaling pathway
• Stimulate variety of responses- blood platelet
aggregation, inflammation, muscle
contraction
• Synthesized from arachidonic acid
 Plant hormones
• Level is regulated by environmental factors in
plant
• Auxins, gibberellins, abscisic acid, cytokinins
and ethylene
• Variety of functions
 Receptors
A receptor is a molecule found on the surface of a cell,
which receives specific chemical signals from neighboring
cells or the wider environment within an organism.

These signals tell a cell to do something—for example to

divide or die, or to allow certain molecules to enter or
exit the cell.

Receptors can be roughly divided into two major classes:

intracellular receptors and extracellular receptors.
EXTRACELLULAR RECEPTORS
• Extracellular receptors are integral
transmembrane proteins and make up most
receptors.
• They span the plasma membrane of the cell, with
one part of the receptor on the outside of the cell
and the other on the inside.
• Signal transduction occurs as a result of a ligand
binding to the outside region of the receptor (the
ligand does not pass through the membrane).
 Intracellular receptors
• receptors located inside the cell rather than on its cell membrane.
• Classic hormones that use intracellular receptors include thyroid
and steroid hormones.
• Examples are:
– Class of nuclear receptors located in the cell nucleus and cytoplasm
– IP3 receptor located on the endoplasmic reticulum.
• The ligands that bind to them are usually:
– Intracellular second messengers like inositol trisphosphate (IP3)
– Extracellular lipophilic hormones like steroid hormones.
• Activated nuclear receptors attach to the DNA at receptor specific
hormone-responsive element (HRE) sequences, located in the promoter
region of the genes activated by the hormone receptor complex.
• Due to their enabling gene transcription, they are alternatively called
inductors of gene expression.
 G-protein coupled receptor
• G protein-coupled receptors are integral membrane proteins
that possess seven membrane-spanning domains or
transmembrane helices.
• These receptors activate a G protein ligand binding.
• G-protein is a trimeric protein. The 3 subunits are called α、β
and γ.
• The α subunit can bind with guanosine diphosphate, GDP. This
causes phosphorylation of the GDP to guanosine
triphosphate, GTP, and activates the α subunit, which then
dissociates from the β and γ subunits.
• The activated α subunit can further affect intracellular
signaling proteins or target functional proteins directly.
 Enzyme-linked receptors
(Protein Tyrosine kinase receptor)
• Enzyme-linked receptors are either enzymes themselves, or are
directly associated with the enzymes that they activate.
• These are usually single-pass transmembrane receptors, with the
enzymatic portion of the receptor being intracellular.
• The majority of enzyme-lined receptors- are protein kinases, or
associate with protein kinases.
• A kinase is a type of enzyme that transfers phosphate groups from
high-energy donor molecules, such as ATP to specific target
molecules (substrates); the process is termed phosphorylation.
• For every phosphorylation event, there is a phosphatase, an
enzyme that can remove phosphate residue and thus modulate
signaling
• Kinase enzymes that specifically phosphorylate tyrosine amino
acids are termed tyrosine kinases . The signal binding domain of
the receptor tyrosine kinase is on the cell surface, while the
tyrosine kinase enzymatic activity resides in the cytoplasmic part of
the protein.
 Cytokine receptors and nonrecptor
protein tyrosine kinase
• Do not have intrinsic enzyme activity
• Stimulate intracellular binding partners
 cAMP pathway: second messengers
and phosphorylation
• It is also known as the adenylyl cyclase pathway.
• In a cAMP-dependent pathway, the activated Gs alpha subunit
binds to and activates an enzyme called adenylyl cyclase , which, in
turn, catalyzes the conversion of ATP into cyclic adenosine
monophosphate (cAMP).
• Increases in concentration of the second messenger cAMP may lead
to the activation of:
• cyclic nucleotide-gated ion channels or an enzyme called protein
kinase (PKA).
• Once PKA is activated, it phosphorylates a number of other proteins
including:
– enzymes that convert glycogen into glucose
– enzymes that promote muscle contraction in the heart leading to an
increase in heart rate
– transcription factors, which regulate gene expression
 cGMP pathway
• Another secondary messenger
• Form via GMP by guanylyl cyclase and
converted back to GMP via GMP
phosphodiesterase
• Role in NO signaling
• Action through protein kinases or ion channels
• Best example conversion of visual signal (light)
to nerve impulse
• Rhodopsin photoreceptor (GPCR)—activated
by light where attached small molecules
isomerizes from 11-cis-retinal to 11-trans-
retinal by absorbing light and induce
conformation change in rhodopsin
• Rhodopsin then activates G protein transducin
• It further stimulates cGMP phosphodiesterase
• It leads to decrease in cGMP conc. Which
affects ion channels
 MAP kinase pathway
• Having cascade of protein kinases
• Highly evolutionary conserved
• Mtiogen-activated-protein kinases (Ser/Thr)
• Activated in response to growth factors and other
signaling molecules
• Controls cell shape , mating cell growth and
proloferation etc.
• Provide a physical link in the signal transduction
pathway from the cytoplasm to the nucleus
• Regulate various transcriptional actions in
Eukaryotic organism. Controls cell growth, to cell
division (mating in haploid yeast).
• ERK (extracellular signal-regulated kinases)-
role in cell proliferation induced by growth
factors activated by many pathways – Ca2+,
protein kinase C or G-PCR (cAMP)
• All intersect at ERK signaling pathway
(stimulating/inhibiting it)
• MAPK also belongs to ERK family
• They act as a relay system of phosphorylation
• ERK activation is mediated by two upstream
kinases which are coupled to growth factor
receptors by Ras GTP-binding protein
• Activation of Ras—activates Raf kinase–
phosphorylateand activate MEK (MAP/ERK
kinase)
• MEK phosphorylate ERK at two positions (Thr
183 & Tyr-185) and activate it
• ERK then phosphorylate its target proteins like
other kinases or transcription factors