BASIC REQUIREMENTS

Cell Signaling
2. The components of signaling pathways
A simple intracellular signaling pathway The first step of the pathway is the binding of the
ligand (first messenger) to the receptor (a G protein-coupled receptor is shown on the slide). The G
protein is the transducer (converter), which affects the first (primary) effector (this is the adenylate
cyclase in the cAMP system, the phospholipase C in the phosphoinositol system). The primary effector
produces the second messenger molecules, which are usually small chemical compounds. The second
messenger is the cAMP itself in the cAMP system, and IP3 and DAG in the phosphoinositol system
(these latter two molecules derive from the PIP 2). The second messenger activates the secondary
effector molecules; these are the protein kinase A in the cAMP system, protein kinase C and Ca 2+ in the
phosphoinositol system. Note that the Ca 2 + is often regarded as a second messenger. The secondary
effector molecule activates further effector molecules (the effect may be negative). The most common
mechanism for activation is the phosphorylation cascade, in which the different kinase molecules
phosphorylate each other in a well-defined sequential order. The protein phosphorylation changes the
conformation of the proteins, which usually means switching to an active state. Various phosphorylase
enzymes carry out the deactivation by removing the phosphate groups. There are hundreds of
different kinase enzyme coding genes in the mammalian genome, which signifies their important role.
The signal transduction cascades change the function of the target proteins which may cause change
in the function of the cells (cellular response). The target proteins could be enzymes of metabolism (it
can change the metabolism), ion channel (it can change the ion milieu), transcription factor (it can
change the gene expression), a cytoskeleton protein (change of the shape and / or movement). The
conditional use is justified because the cell is a “decision machine” that can override the effect of a
signal (another effect of signals), or neglect the command, because intensity of the signal is too low.
The cell responses are usually dependent on signal intensity; in case of stronger signal, the response is
greater. The incoming input (neurotransmitters) of the neurons exerts a gradual response in the body
of the cell (continuous ion gradient), but the output signal (action potential) is discrete, in particular, is
always the same size, and therefore comparable to digital (yes or no basis). The signal intensity of
neurons is encoded by the frequency of action potentials (and not the strength of individual action
potential).

SIGNAL MOLECUES (ligands) responsible for transmission of the information between cells.
The signaling molecules are produced by signaling cells, they have to move through the gap between
the cells in order to bind to the receptor of the receiving cells and evoke a response in the target cells.
The main types of the signaling molecules are as follows: hormones, growth and differentiation
factors, chemokines, cytokines, neurotransmitters, nitrogen oxide (NO), etc. Molecules similar to the
signal molecules of the organism can be found in nature too, and we can produce them synthetically,
too. Caffeine, nicotine and certain drugs are similar to certain naturally occurring ligands and
therefore, they can be recognized by the appropriate receptors. Some drugs also bind to our
receptors. With these substances, we can control cell communication and bodily functions.

Cytokines are small protein or peptide molecules that are secreted by the glial cells and by numerous cells of the
immune system, and are a category of signaling molecules used in intercellular communication. As we learn
more about them, the distinctions between cytokines and hormones are fading. Cytokines are not limited to
their immunomodulatory role; these molecules are also involved in several developmental processes during
embryogenesis. The effect of a particular cytokine on a given cell depends on its extracellular abundance, the
presence and abundance of their receptors on the cell surface, and downstream signals activated by receptor
binding; these last two factors can vary by cell type. Cytokines have been classed as lymphokines, interleukins,
and chemokines, based on their presumed function, cell of secretion, or target of action. Cytokines are

CELL SIGNALING– II. The components of signaling pathways page 1

 BASIC REQUIREMENTS

characterized by considerable redundancy (many cytokines share similar functions) and pleiotropism (a single
cytokine affects a number of processes). Cytokines may be divided into six groups: interleukins, colony-
stimulating factors, interferons, tumor necrosis factor, growth factors, and chemokines. The term chemokine
refers to a specific class of cytokines that mediates chemo-attraction between cells.

A growth factor is a naturally occurring substance capable of stimulating cellular growth, proliferation and
cellular differentiation. Usually it is a protein or a steroid hormone. Growth factors are important for regulating a
variety of cellular processes. They often promote cell differentiation and maturation, which varies between
growth factors. For example, bone morphogenic proteins (BMPs) stimulate bone cell differentiation, while
fibroblast growth factors (FGFs) and vascular endothelial growth factors (VEGFs) stimulate blood vessel
differentiation (angiogenesis). Growth factor is sometimes used interchangeably among scientists with the term
cytokine. While growth factor implies a positive effect on cell division, cytokine is a neutral term with respect to
whether a molecule affects proliferation. While some cytokines can be growth factors, such as granulocyte
colony-stimulating factor (G-CSF), others have an inhibitory effect on cell growth or proliferation. Some
cytokines, such as Fas-ligand, are used as "death" signals; they cause target cells to undergo programmed cell
death (apoptosis).

Foreign signals Beside endogenous ligands, many signaling pathways can be targeted by natural or
synthetic exogenous substances, such as snake venom, drugs, nicotine, kaffein, products of
pharmaceutical industries, etc.

RECEPTORS are responsible for the perception of signals and converting them into other types of
signals, which then are recognized by the signal processing apparatus of the cell eliciting a response.
The relationship between receptors and ligands is characterized by the key – lock relationship, the
steric configuration/shape of the receptor allows only the ligand that fits into this structure to bind
(the recognition site). The three-dimensional "complementarity" is not enough, the two molecules
should be able to establish chemical bond formation (e.g.: H-bonds, ionic bonds, etc). The receptors
can be categorized on the basis of their position in the cell. Thus we talk about cytoplasmic (nuclear)
and cell-surface receptors. The steroid hormones are typical ligands of cytoplasmic receptors, are
relatively low in molecular weight lipophilic (fat-soluble) molecules can easily penetrate through the
cell membrane, bind to the receptor, the receptor-ligand complex becomes active as transcription
factor . The receptor-ligand complex crosses the nuclear membrane and binds to the recognition site
of its promoter or enhancer DNA sequences, and thereby stimulates the expression of corresponding
genes. The cell surface receptors have three main types, which are ionotropic (ion channel-coupled),
G-protein-linked, and enzyme-coupled receptors.

1. Signal pathways without receptors

Nitric oxide (NO) Nitric oxide (NO) acts by a direct activation on an enzyme. Acetyl-choline released
by nerve terminals in the blood vessel wall stimulates endothelial cells lining the blood vessel to make
and release NO. No is made from the amino acid arginine by the enzyme NO synthase. The gas has a
very short half-life (about 5-10 seconds) because it is quickly converted to nitrates and nitrites by
cellular enzymes. The NO diffuses out of the endothelial cells and into adjacent smooth muscle cells,
causing the muscle cells to relax, allowing the vessel to dilate, so that blood flows through it more
freely. The effects of NO on blood vessels accounts for the action of nitroglycerine (it is converted to
NO in the body), which has been used for almost 100 years to treat patients with angina (pain caused
by inadequate blood flow to the heart muscle from the coronary blood vessels). Inside many target
cells, NO binds to the enzyme guanylate cyclase stimulating the formation of cyclic GMP from the
nucleotide GTP. Cyclic GMP itself is a small intracellular signaling molecule that forms the next link in
the signaling chain that leads to the cell’s ultimate response. The impotence drug Viagra enhances
penis erection by blocking the degradation of cyclic GMP, prolonging the NO signal.

CELL SIGNALING– II. The components of signaling pathways page 2

 BASIC REQUIREMENTS

2. Intracellular receptor-mediated signaling pathways

Hydrophobic signal molecules such as the steroid hormones – including cortisol (glucocorticoid),
estradiol and testosterone (sex steroids) – and the thyroid hormones such as thyroxin all pass through
the plasma membrane of the target cells, where they bind to an intracellular receptor. The
intracellular receptor is in fact, an inactive transcription factor, which is activated upon binding to the
steroid hormone. Steroid hormone receptors play an essential role in human physiology, as illustrated
by the dramatic consequences of a lack of the receptor for testosterone in humans. These individuals
are genetically males (they have Y chromosomes) but they develop as females.

An example: Glucocorticoid receptors are located in the cytoplasm in an inactive state ( hsp90 chaperons
inhibit them). Steroid hormone binding dislocates hsp90, and results in the formation of a dimeric ( two-
subunit) molecule, which in turn, enter to the nucleus, bind to its response DNA element ( GRE:
glucocorticoid response element) and activate transcription from the linked gene.

(1) Steroid hormones, which include the cortisol, the steroid sex hormones and vitamin D, are all made from
cholesterol. (a) Cortisol is produced in the cortex of the adrenal gland and influences the metabolism of many cell
types. (b) The steroid sex hormones are made in the testis and ovaries, and they are responsible for the primary
and secondary sex characteristics that distinguish males from females. (c) Vitamin D is synthesized in the skin in
response to sunlight; after it has converted to its active form in the liver or kidneys, it regulated Ca 2+ metabolism.
(2) The thyroid hormones, which are made from the amino acid tyrosine, act to increase the metabolic rate in
many cell types. (3) Retinoids, such as retinoic acid, are made from vitamin A, and have important role as local
mediators in vertebrate development.

Nuclear receptors Some of the nuclear receptors, such as those for cortisol, are located primarily in the cytosol
and enter the nucleus only after ligand binding; others, such as thyroid and retionid receptors, are bound to the
DNA in the absence of ligand. In either case the inactive receptors are usually bound to inhibitory protein
complexes. Ligand binding alters the conformation of the receptor protein, causing the inhibitory complex to
dissociate, while also causing the receptor bind co-activator proteins that stimulate gene transcription. In other
cases, however, ligand binding to a nuclear receptor inhibits transcription: some thyroid hormone receptors, for
example, act as transcriptional activators in the absence of their hormones and become transcriptional
repressors when hormone binds. The transcriptional response usually takes place in multiple steps. Some of the
primary-response proteins turn on secondary-response genes, whereas others turn off the primary-response
genes. The intracellular receptors are all structurally related, being part of the very large nuclear receptor
superfamily. Many family members are so called orphan receptors, indicating that their ligands are unknown.

3. Cell surface receptor-mediated signaling pathways

The vast majority of signal molecules are too large or hydrophilic to cross the plasma membrane of the
target cell. This bulky molecules bind to receptor proteins that span the plasma membrane. These
receptors detect a signal on the outside and relay the message, in a new form, into the interior of the
cell. The cell surface receptors belong to one of the three large families: (1) ion-channel linked
receptors; (2) G protein-linked receptors, or (3) enzyme-linked receptors. The number of different
types of receptors is greater than the extracellular signal that act on them, because for many
extracellular signal molecules there is more than one type of receptor. The neurotransmitter
acetylcholine, for example, acts on skeletal muscle cells via an ion-channel-linked receptor, whereas in
heart muscle it acts through G-protein linked receptor. These two types of receptors generate
different intracellular signals, and thus enable the two types of muscles to react to acetylcholine in
different ways, increasing contraction in skeletal muscle and decreasing the frequency of contractions
in heart. Many foreign targets can interfere with our physiology. These substances either mimic the
natural ligand for a receptor, occupying the normal ligand-binding site, or bind to the receptor at some
other site, blocking or over-stimulating the receptor’s natural activity. Many drugs and poisons act in
this way.

CELL SIGNALING– II. The components of signaling pathways page 3

 BASIC REQUIREMENTS

3a. Ion channel-coupled receptors (in other names: ionotropic receptors, or ligand-gated ion channels,
neurotransmitter-gated ion channels). When a neurotransmitter binds, this type of receptor alters its
conformation so as to open or close a channel for the flow of specific types of ions - such as Na +, K+,
Ca2+, or Cl- - across the plasma membrane. Driven by electrochemical gradient, the ions rush into or out
of the cell, creating a change in the membrane potential within a millisecond, or so. This change in
potential may trigger a nerve impulse, or alter the ability of other signals to do so. Whereas ion-
channel-linked receptors are specialty of the nervous system and other electrically excitable cells such
as muscle, G-protein linked receptors and enzyme-linked receptors are used by practically every cell
type of the body. See this topic in more detail in “Neural communication” lecture.

3b. G protein-coupled receptors (GPCRs) G-protein-linked receptors form the largest family of
receptors, with hundreds of members already identified in mammalian cells. They mediate most
responses to signals from the external world, as well as signals from other cells, including hormones,
neurotransmitters, and local mediators. There are more than 800 GPCRs in humans, and in mice there
are about 1000 concerned with the sense alone (in mice receptor genes were generated by gene
duplications followed by functional divergence, while these genes were lost and mutated in the human
genome). The signal molecules that act on GPCRs include proteins and small polypeptides, as well as
derivatives of amino acids and fatty acids, not to mention photons of light and all the molecules that
we can smell or taste. The same signal molecule can activate many different GPCR family members; for
example, adrenaline activates at least 9 distinct GPCRs, acetyl choline another 5, and serotonin at least
14. The different receptors for the same signal are usually expressed in different cell types and elicit
different responses.

GPCR kinases (GRK; G protein-coupled receptor kinase) A GRK phosphorylates only activated
receptors. The binding of an arrestin to the phosphorylated receptor prevents the receptor from
binding to its G protein. GRK-mediated inactivation of GPCRs is a mechanism act to desensitization of
the receptor in case of prolonged exposition to a high concentration of ligand molecules.

G proteins (trimeric G proteins) When an extracellular signal molecule binds to a GPCR, the receptor
undergoes a conformational change that enables it to activate a G protein. In some cases, the G
protein is physically associated with the receptor before the receptor is activated, whereas in others it
binds only after receptor activation. There are various types of G proteins, each specific for a particular
set of GPCRs and for a particular set of target proteins in the plasma membranes. G proteins are
composed of three subunits – ,  and . In the unstimulated state, the  subunit has GDP bound and
the G protein is inactive. When a GPCR is activated, it acts like a guanine nucleotide exchange factor
(GEF) and induces the  subunit to release its bound GDP, allowing GTP to bind in its place. When a
ligand binds to its receptor, the altered receptor activates the G protein by causing the  subunit to
lose its affinity to GDP, which it exchanges for a molecule of GTP. The activated GTP-bound  subunit
then dissociate from the  complex (thereby activating them). The two activated parts of a G protein
- the  subunit and  complex - then interact with their targets to elicit intracellular response. The
amount of time that the two parts of G protein remain dissociated, and - hence available to relay
signals – is limited by the intrinsic GTPase activity of the  subunit. The reassociation generally occurs
within seconds after the G protein has been activated. The cholera toxin enters the cells that lines the
intestine and modifies the  subunit in such a way that it can no longer hydrolyze its bound GTP,
thereby the  subunit remains in the active state indefinitely. This results in excessive outflow of water
into the gut, and thus dehydration.

3c. Enzyme-linked receptors are the receptors for many growth factors, cytokines and hormones. The
cytoplasmic domain of enzyme –linked receptors act as enzymes – or form complexes with other
proteins act as enzymes. These receptors came to light through their role in responses to extracellular
signal proteins that regulate growth, proliferation, differentiation, and survival of cells. Most of these
proteins act as local mediators and act at very low concentrations. Mutations in receptor tyrosine

CELL SIGNALING– II. The components of signaling pathways page 4

 BASIC REQUIREMENTS

kinases are responsible for a wide array of diseases, including cancers, neurodegeneration,
achondroplasia and atherosclerosis.
There are five main types of enzyme-linked receptors:
1. Receptor tyrosine kinase (RTK): Contains intrinsic tyrosine kinase activity (EGFR, VEGFR)
2. Receptor serine/threonine Kinase: Contains intrinsic serine/threonine kinase activity (TGF-βR)
3. Receptor guanylate cyclase: Contain intrinsic cyclase activity (ANP)
4. Tyrosine-kinase associated receptors: Receptors that associate with proteins that have tyrosine
kinase activity (cytokine receptors)
5. Receptor tyrosine phosphatases

The largest class of enzyme-linked receptors made up of those with a cytoplasmic domain that
functions as a tyrosine protein kinase, phosphorylating tyrosine side chains on selected intracellular
proteins. Such receptors are called receptor tyrosine kinases. The binding of a signal molecule to the
extracellular domain of a receptor tyrosine kinase causes two receptor molecules to associate into a
dimer. The signal molecule (as shown in our example) is itself a dimer and thus can physically cross-link
the two receptors. In other cases, binding the signal molecule changes the conformation of the
receptor molecules in such a way that they dimerize. Dimer formation brings the kinase domains of
each intracellular receptor tail into contact with the other; this activates the kinases and enables them
to phosphorylate each other at several tyrosine side chains. Each phosphorylated tyrosine serves as a
specific site for different (as many as 10 or 20) intracellular signaling proteins, which in turn, transmit
their signals along several routes simultaneously to many destinations inside the cell, thus activating
and coordinating the numerous biochemical changes, that are required to trigger a complex response,
such as cell proliferation. To terminate the activation of the receptor, the cell contains protein tyrosine
phosphatase enzymes, which remove the phosphates from the receptor (see below for other
strategies of receptor inactivation). Different receptor tyrosine kinases recruit different collections of
intracellular signaling proteins, producing different effects; but certain components are used quite
widely. The main signaling pathway from receptor tyrosine kinases to the nucleus is the MAP-kinase
pathway.

Monomeric G proteins (monomeric GTPases) The most representative members of monomeric G

proteins are the Ras proteins. There are three major, closely related Ras proteins in humans (H-, K-,
and N-Ras). Ras contains one or more covalently attached lipid groups that help anchor the protein to
the cytoplasmic face of the membrane. Ras functions as a molecular switch, cycling between two
distinct conformational states – active when GTP is bound and inactive when GDP is bound.

Intracellular signaling molecules

(I) The small intracellular mediators, or second messengers, are generated in large numbers in
response to receptor activation and often diffuse away from their source, spreading the signal to other
parts of the cell. In addition to their job as relay molecules, second messengers serve to greatly amplify
the strength of the signal. Binding of a ligand to a single receptor at the cell surface may end up
causing massive changes in the biochemical activities within the cell. Some, such as cyclic AMP and
Ca2+, are water soluble and diffuse in the cytosol, while others, such as diacylglycerol (DAG), are lipid-
soluble and diffuse in the plane of the plasma membrane. In either case, they pass the signal on by
binding to and altering the conformation and behavior of selected signaling proteins or effector
proteins. There are 4 major classes of second messengers:
1. cyclic nucleotides (e.g., cAMP and cGMP)
2. inositol triphosphate (IP3)and diacylglycerol (DAG)
3. calcium ions (Ca2+)
4. gases (NO, CO)

(II) Intracellular signaling proteins help relay the signal into the cell by either generating second
messenger molecules or activating the next signaling or effector protein in the pathway. These

CELL SIGNALING– II. The components of signaling pathways page 5

 BASIC REQUIREMENTS

proteins form a functional network, in which each protein helps to process the signal in one or more of
the following ways as it spreads the signal’s influence through the cell. (1) The protein may simply
relay the signal to the next signaling component in the chain. (2) It may act as a scaffold to bring two or
more signaling proteins together so that they can interact more quickly and efficiently. (3) It may
transform (or transduce) the signal into different form, which is suitable for either passing the signal,
or along or stimulating a cell response. (4) It may amplify the signal it receives, either by producing a
large amount of small second messenger molecules, or by activating many copies of a down-stream
signaling molecule. When there are multiple amplification steps in a relay chain, the chain is often
referred to as a signaling cascade. (5) It may receive signals from two or more signaling pathways and
integrate them before relaying a signal onward. A protein that that requires input from two or more
signaling pathways to become activated, is often referred to as a coincidence detector. 6) It may
spread the signal from one signaling pathway to another, creating branches in the signaling stream,
thereby increasing the complexity of the response. (7) It may anchor one or more signaling proteins in
a pathway to a particular structure in the cell where the signaling proteins are needed. (8) It may
modulate the activity of other signaling proteins and thereby regulating the strength of signaling along
a pathway.

Activation and inhibition of the components of signaling pathways Many intracellular signaling
proteins behave like molecular switches. When they receive a signal, they switch from an inactive to
an active conformation, until another process switches them off. Two important classes of molecular
switches that operate in intracellular signaling pathways depend on the gain or loss of phosphate
groups for their activation or inactivation, although the way in which the phosphate is gained and lost
is very different in the two classes. One of these processes is the phosphorylation (or
dephosphorylation), and the other one is the exchange of GDP to GTP, or vice versa. The modification
of the activity of signal proteins can occur in other ways too (see below).

1. Phosphorylation
2. GTP-binding
3. Binding to a second messenger
4. Proteolytic cleavage
5. Dissociation of an inhibitory protein (chaperon)
6. Ubiquitination

Protein kinases: protein phosphorylation The largest class of signaling proteins are activated or
inactivated by phosphorylation. The activity of any protein regulated by phosphorylation depends on
the balance between the activities of the kinases that phosphorylate it and phosphatases that
dephosphorylate it. About 30% of human proteins contain covalently attached phosphate, and the
human genome encodes about 520 protein kinases and about 150 protein phosphatases. Many
signaling proteins controlled by phosphorylation are themselves protein kinases, and these are often
organized into phosphorylation cascades. In such a cascade, one protein kinase, activated by
phosphorylation, phosphorylates the next protein kinase in the sequence, and so on, relaying the
signal onward. The great majority of these enzymes are (1) serine/threonin kinases, which
phosphorylate proteins on serines and less often on threonines. (2) Others are tyrosine kinases, which
phosphorylate proteins on tyrosines.

GTP binding proteins: The other important class of molecular switches that function by gaining and
losing phosphate groups consists of GTP-binding proteins. These proteins switch between an “ON”
state when GTP is bound and an “OFF” state when GDP is bound. In the “ON” state they have intrinsic
GTPase activity and shut themselves off by hydrolyzing their GTP to GDP. There are two major types of
GTP-binding proteins. (1) Large trimeric GTP-binding proteins (also called G proteins) help relay signal
from G-protein-coupled receptors that activate them. (2) Small monomeric GTP-binding proteins (also
called monomeric GTPases) help relay signals from many classes of cell-surface receptors. GTPase

CELL SIGNALING– II. The components of signaling pathways page 6

 BASIC REQUIREMENTS

activator proteins (GAPs) drive the proteins into an “OFF” state by hydrolyzing the bound GTP to GDP.
Conversely, G-protein-coupled receptors activate trimeric G proteins, and guanine nucleotide
exchange factors (GEFs) activate monomeric GTPases, by promoting the release of bound GDP in
exchange for binding of GTP (no phosphorylation of GDP to GTP in a bound state!). Not all signaling
proteins act as switches when they are phosphorylated or otherwise modified, covalently added
groups can simply mark the protein so that it can interact with other signaling proteins that recognize
the modification.

Other switches Not all molecular switches in signaling pathways depend on phosphorylation or GTP
binding, however. Some signaling proteins are switched on or off by binding of another signaling
proteins (e.g. inhibitors, such as chaperons), or a small intracellular mediator (second messenger) such
as cAMP or Ca2+, or by covalent modifications other than phosphorylation, such as ubiquitylation.
Proteolytic cleavage (such as in caspases) is another form of activation.

Inhibitors In many signal pathways, beside the lack of incoming signal, the inactivity of a component is
further ensured by binding an inhibitory protein.

Proteolysis „chain reaction” In the process of apoptosis the proteases called caspases activate each
other by cleaving a short peptide from the pro-caspase molecules. The so generated active caspase
does the same with another procaspase molecule, and so on (caspase cascade).

The role of calcium in the cell Calcium ions are one of the most widespread second messengers used
in signal transduction. They make their entrance into the cytoplasm either from outside the cell
through the cell membrane via calcium channels, or from the internal calcium storages such as
endoplasmic reticulum or mitochondria. The level of intracellular calcium is regulated by transport
proteins. The sodium-calcium exchanger uses energy from the electrochemical gradient of Na + pumps
out Ca2+ from the cell in exchange for the entry of Na +. Additionally, Ca2+ pumps obtain energy to
remove calcium from the cell by the hydrolysis of ATP. In neurons, voltage dependent calcium-
selective ion channels are important for synaptic transmission through triggering the release of
neurotransmitters through the fusion of neurotransmitter vesicles with the plasma membrane of axon
terminal of post-synaptic (sending) neuron.

CAM-kinase-II An increase of the level of free Ca 2+ in the cytosol is triggered by many different signals.
Ca2+ exerts its effect by binding to and influencing the activity of Ca 2+ –binding proteins. The most
widespread and common of these is the calmodulin, which is present in the cytosol of all eukaryotic
cells. When calmodulin binds to Ca 2+, the protein undergoes conformational change that enables it to
wrap around a wide range of target proteins, altering their activities. One particularly important class
of targets for calmodulin is the Ca 2+ /calmodulin-dependent kinases (CaM-kinases). When these
kinases are activated by binding to calmodulin complexed with Ca 2+, they influence other processes in
the cell by the phosphorylation of selected proteins.

Chapter 15 Cell signaling and communication in the textbook LIFE is only a facultative material, but it
helps understanding the topic, therefore, it is highly recommended to read.

http://www.bio.davidson.edu/courses/Immunology/Flash/IP3.html

CELL SIGNALING– II. The components of signaling pathways page 7