Signal Transduction

cGMP as secondary messenger

• Cyclic guanosine monophosphate (cGMP) is a cyclic nucleotide derived from guanosine triphosphate (GTP). cGMP acts as
a second messenger much like cyclic AMP. Its most likely mechanism of action is activation of intracellular protein kinases in
response to the binding of membrane-impermeable peptide hormones to the external cell surface.
• Functions of cGMP
• cGMP is a common regulator of ion channel conductance, glycogenolysis, and cellular apoptosis. It also relaxes smooth
muscle tissues. In blood vessels, relaxation of vascular smooth muscles lead to vasodilation and increased blood flow.
• cGMP is a secondary messenger in phototransduction in the eye. In the photoreceptors of the mammalian eye, the presence
of light activates phosphodiesterase, which degrades cGMP. The sodium ion channels in photoreceptors are cGMP-gated, so
degradation of cGMP causes sodium channels to close, which leads to the hyperpolarization of the photoreceptor's plasma
membrane and ultimately to visual information being sent to the brain.
• cGMP is also seen to mediate the switching on of the attraction of apical dendrites of pyramidal cells in cortical layer
V towards semaphorin-3A (Sema3a). Whereas the axons of pyramidal cells are repelled by Sema3a, the apical dendrites are
attracted to it. The attraction is mediated by the increased levels of soluble guanylate cyclase (SGC) that are present in the
apical dendrites. SGC generates cGMP, leading to a sequence of chemical activations that result in the attraction towards
Sema3a. The absence of SGC in the axon causes the repulsion from Sema3a. This strategy ensures the structural polarization
of pyramidal neurons and takes place in embryonic development.
• cGMP, like cAMP, gets synthesized when olfactory receptors receive odorous input. cGMP is produced slowly and has a more
sustained life than cAMP, which has implicated it in long-term cellular responses to odor stimulation, . cGMP in the olfactory
is synthesized by both membrane guanylyl cyclase (mGC) as well as soluble guanylyl cyclase (sGC). Studies have found that
cGMP synthesis in the olfactory is due to sGC activation by nitric oxide, a neurotransmitter. cGMP also requires increased
intracellular levels of cAMP and the link between the two second messengers appears to be due to rising intracellular
calcium levels.
• cGMP is involved in the regulation of some protein-dependent kinases. For example, PKG (protein kinase G) is
a dimer consisting of one catalytic and one regulatory unit, with the regulatory units blocking the active sites of the catalytic
units. cGMP binds to sites on the regulatory units of PKG and activates the catalytic units, enabling them to phosphorylate
their substrates.
 “NO” as intercellular secondary messenger

• During the 1980s, a new type of messenger was discovered that was neither an organic compound, such
as cAMP, nor an ion, such as Ca 2+ ; it was an inorganic gas—nitric oxide ( NO ). NO is unusual because it
acts both as an extracellular messenger, mediating intercellular communication, and as a second
messenger, acting within the cell in which it is generated. NO is formed from the amino acid L ‐arginine in
a reaction that requires oxygen and NADPH and that is catalyzed by the enzyme nitric oxide synthase (
NOS ). Since its discovery, it has become evident that NO is involved in a myriad of biological processes
including anticoagulation, neurotransmission, smooth muscle relaxation, and visual perception.

• The binding of acetylcholine to the outer surface of an endothelial cell signals a rise in cytosolic Ca 2+
concentration that activates nitric oxide synthase . The NO formed in the endothelial cell diffuses across
the plasma membrane and into the adjacent smooth muscle cells , where it binds and stimulates guanylyl
cyclase , the enzyme that synthesizes cyclic GMP (cGMP), which is an important second messenger . Cyclic
GMP binds to a cGMP‐dependent protein kinase (a PKG), which phosphorylates specific substrates causing
relaxation of the muscle cell and dilation of the blood vessel.
 Contd…

• Nitroglycerine, which had been used since the 1860s to treat the painof angina that results from an
inadequate flow of blood to the heart.Ni troglycerine is metabolized to nitric oxide, which stimulates the
relaxation of the smooth muscles lining the blood vessels of the heart, increasing blood flow to the organ.
The therapeutic benefits of nitroglycerine were discovered through an interesting observation.Persons
with heart disease who worked with nitroglycerine in Alfred Nobel ’ s dynamite factory were found to
suffer more from the pain of angina on days they weren ’ t at work. It is only fitting that the Nobel Prize,
which is funded by a donation from Alfred Nobel ’ s estate, wasawarded in 1998 for the discovery of NO as
a signaling agent.
• Recent investigations have revealed that NO has a variety of actions within the body that do not involve
production of cGMP.For example, NO is added to the — SH group of certain cysteine residues in well over
a hundred proteins, including hemoglobin, Ras, ryanodine channels, and caspases. This posttranslational
modification, which is called S‐nitrosylation , alters the activity, turnover, or interactions of the protein.
Calcium (Ca+2 ) as an intracellular secondary messenger

• Calcium ions play a significant role in a remarkable variety of cellular activities, including muscle
contraction, immune responses, cell division, secretion, fertilization, synaptic transmission,
metabolism, transcription, cell movement, and cell death.
• In each of these cases, an extracellular message is received at the cell surface and leads to a
dramatic increase in concentration of calcium ions within the cytosol.
• The concentration of calcium ions in a particular cellular compartment is controlled by the
regulated activity of Ca 2+ pumps, Ca 2+ exchangers, and/or Ca 2+ ion channels located within the
membranes that surround the compartment (as in Figure 15.28 ).
• The concentration of Ca 2+ ions in the cytosol of a resting cell is maintained at very low levels,
typically about 10-7M. In contrast, the concentration of this ion in the extracellular space or within
the lumen of the ER or a plant cell vacuole is typically 10,000 times higher than the cytosol. The
cytosolic calcium level is kept very low because (1) Ca 2+ ion channels in both the plasma and ER
membranes are normally kept closed, making these membranes highly impermeable to this ion,
and (2) energy‐driven Ca 2+ transport systems of the plasma and ER membranes pump calcium out
of the cytosol. 2 Abnormal elevation of cytosolic Ca 2+ concentration, as can occur in brain cells
followinga stroke, can lead to massive cell death.
• Extracellular signals that are transmitted by Ca 2+ ions typically act by opening a small number of Ca 2+
ion channels at the cell surface at the site of the stimulus. As Ca 2+ ions rush through these channels and
enter the cytosol, they act on nearby Ca 2+ ion channels in the ER, causing these channels to open and
release additional calcium into adjacent regions of the cytosol. In some responses, the elevation of Ca 2+
levels remains localized to a small region of the cytosol . In other cases, a propagated wave of calcium
release spreads through the entire cytoplasmic compartment.
 Contd….

• Unlike cAMP, whose action is usually mediated by stimulation of a protein kinase, calcium can affect a
number of different types of cellular effectors, including protein kinases .
• Depending on the cell type, calcium ions can activate or inhibit various enzyme and transport systems,
change the ionic permeability of membranes, induce membrane fusion, or alter cytoskeletal structure and
function.
• Calcium does not bring about these responses by itself but acts in conjunction with a number of
calcium‐binding proteins. The best‐studied calcium‐binding protein is calmodulin , which participates in
many signaling pathways.
• Calmodulin is found universally in plants, animals, and eukaryotic microorganisms, and it has virtually the
same amino acid sequence from one end of the eukaryotic spectrum to the other.
• Each molecule of calmodulin contains four binding sites for calcium. Calmodulin does not have sufficient
affinity for Ca 2+ to bind the ion in a nonstimulated cell. If, however, the Ca 2+ concentration rises in
response to a stimulus, the ions bind to calmodulin, changing the conformation of the protein and
increasing its affinity for a variety of effectors. Depending on the cell type, the calcium–calmodulin (Ca 2+
–CaM) complex may bind to a protein kinase, a cyclic nucleotide phosphodiesterase, ion channels, or even
to the calcium‐transport system of the plasma membrane.
• Regulating Calcium Concentrations in Plant Cells-
• Calcium ions (acting in conjunction with calmodulin) are important intracellular messengers in plant
cells. The levels of cytosolic calcium change dramatically within certain plant cells in response to a
variety of stimuli, including changes in light, pressure, gravity, and
• the concentration of plant hormones such as abscisic acid. The concentration of Ca 2+ in the cytosol
of a resting plant cell is kept very low by the action of transport proteins situated in the plasma
membrane and vacuolar membrane (tonoplast).
• The role of Ca 2+ in plant cell signaling is illustrated by guard cells that regulate the diameter of the
microscopic pores (stomata) of a leaf . Stomata are a major site of water loss in plants, and the
diameter of their aperture is tightly controlled, which prevents dehydration. The diameter of the
stomatal pore decreases as the fluid (turgor) pressure in the guard cell decreases. The drop in
turgor pressure is caused in turn by a decrease in the ionic concentration (osmolarity) of the guard
cell. Adverse conditions, such as high temperatures and low humidity, stimulate the release of the
plant stress hormone abscisic acid. Studies suggest that abscisic acid binds to a GPCR in the plasma
membrane of guard cells, triggering the opening of Ca 2+ ion channels in the same membrane .
The resulting influx of Ca 2+ into the cytosol triggers the release of additional Ca 2+ from
intracellular stores. The elevated cytosolic Ca 2+ concentration leads to closure of K + influx
channels in the plasma membrane and opening of bothK + and anion efflux channels. These
changes produce a net outflow of K + ions and anions ( NO 3 − and Cl − ) and a resulting decrease in
turgor pressure.
Contd…