Pharmaceutical Biochemistry II (Theory) Lecture 17 (Spring-2016)

Second messengers and their role in the Regulation of Metabolic Processes

Second messenger
These are intracellular chemical signals with short half-life. The concentration of these
signals is regulated by different hormones, neurotransmitters, and other extracellular signals.
Second messengers include cAMP, cGMP, Ca2+, Inositol Triphosphate, Diacylglycerol and
Nitrogen Monoxide.

A. Cyclic AMP (cAMP or 3'-5'-cyclic adenosine monophosphate)

Cyclic AMP is synthesized by action of an enzyme adenylate cyclase. This enzyme cyclizes
the ATP to cAMP by releasing pyrophosphate (PPi) molecule. The degradation of cAMP to
AMP is catalyzed by phosphodiesterases. The enzyme adenylate cyclase activity is regulated
by G proteins and activities of G proteins are controlled by extracellular signals from 7-helix
receptors (β-adrenergic receptor in plasma membrane).

The 7-Helix receptors [also called as serpentine receptors, G protein-coupled receptors

(GPCR), or 7 transmembrane segment (7tm) receptors] represent a large group of membrane
proteins that transfer the hormone or transmitter signal, with the help of G proteins to the
effecter proteins that alter the concentrations of ions and second messengers.

G proteins consist of three different types of subunit (α, β, and γ). The α-subunit can bind
GDP or GTP (hence the name “G protein”). G proteins are divided into several types,
depending on their effects. Stimulatory G proteins (Gs) activate adenylate cyclases or
influence ion channels. Inhibitory G proteins (Gi) inhibit adenylate cyclase. G proteins in the
Gq family activate another effector enzyme, phospholipase c.

The cAMP is a second messenger, used for intracellular signal transduction, such as
transferring the effects of hormones like glucagon and adrenaline, which cannot get through
the cell membrane. It is involved in the activation of protein kinases and regulates the effects
of adrenaline and glucagon. It also regulates the passage of Ca 2+ through ion channels. The
cAMP and its associated kinases function in several biochemical processes, including the
regulation of glycogen, sugar, and lipid metabolism.

In humans, cyclic AMP works by activating protein kinase A (PKA). The PKA is normally an
inactive tetrameric holoenzyme, consisting of two catalytic and two regulatory units (C 2R2).

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Pharmaceutical Biochemistry II (Theory) Lecture 17 (Spring-2016)

The regulatory units keep the catalytic centers of the catalytic units blocked. This process is
called as auto-inhibition. Cyclic AMP binds to specific locations on the regulatory units of the
protein kinase, and causes dissociation between the regulatory and catalytic subunits, thus
activating the catalytic units and enabling them to phosphorylate the substrate proteins. The
active subunits catalyze the transfer of phosphate from ATP to specific serine or threonine
residues of protein substrates. Phosphorylated proteins may act directly on the cell ion
channels. The PKA can also phosphorylate specific proteins that bind to promoter regions of
DNA, causing increased expression of specific genes.

Not all protein kinases respond to cAMP. Several classes of protein kinases, including protein
kinase C, are not cAMP-dependent. Further effects mainly depend on protein kinase, which
vary on the bases of type of cell. There are some minor PKA-independent functions of cAMP,
e.g. activation of calcium channels, providing a minor pathway by which growth hormone
releasing hormone causes release growth hormone.

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Pharmaceutical Biochemistry II (Theory) Lecture 17 (Spring-2016)

Figure 12-16 is only for knowledge and understanding, not for examination

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Pharmaceutical Biochemistry II (Theory) Lecture 17 (Spring-2016)

B. Phosphoinositol
Phosphoinositol cascade also converts extracellular signals into intracellular ones. The
intracellular messengers formed by activation of this pathway arise by cleavage of
phosphatidyl-inositol 4,5-bisphosphate (PIP2). The binding of a hormone such as vasopressin
to a 7-Helix receptor leads to the activation of the isoform of phospholipase C. The G
protein that activates phospholipase C is called Gq. The activated enzyme then hydrolyzes the
phosphodiester bond linking the phosphorylated inositol unit to the acylated glycerol moiety.
The cleavage of PIP2 produces two messengers: inositol 1,4,5-trisphosphate (InsP3), a soluble
molecule that can diffuse from the membrane, and diacylglycerol, which stays in the
membrane. InsP3 migrates to endoplasmic reticulum and opens Ca2+ channels that allow Ca2+
to flow into cytoplasm. The elevated level of Ca2+ in the cytosol then triggers processes such
as smooth muscle contraction, glycogen breakdown, and vesicle release. Diacylglycerol,
which is lipophilic, remains in the membrane, where it activates protein kinases C, which
phosphorylate proteins in the presence of Ca2+ ions and thereby pass the signal.

C. Calcium ions
Ca2+ is a signaling substance. The concentration of Ca2+ ions in the cytoplasm is normally
very low (10-100 nM), as it is kept down by ATP driven Ca 2+ pumps and by Na+/Ca2+
exchangers. In addition, many proteins in the cytoplasm and organelles bind calcium and thus
act as Ca2+ buffers. Specific signals e.g. an action potential or second messenger such as InsP3
or cAMP, can trigger a sudden increase in the cytoplasmic Ca 2+ level to 500-1000 nM by
opening Ca2+ channels in the plasma membrane or in the membranes of the endoplasmic or
sarcoplasmic reticulum. In the cytoplasm, the Ca2+ level always only rises very briefly, as

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Pharmaceutical Biochemistry II (Theory) Lecture 17 (Spring-2016)

prolonged high concentrations in the cytoplasm have cytotoxic effects. The biochemical
effects of Ca2+ in the cytoplasm are mediated by special Ca2+-binding proteins. These include
the annexins, calmodulin, and troponin C in muscle. Calmodulin is a relatively small protein
(17 kDa) that occurs in all animal cells. Binding of four Ca 2+ ions converts it into a regulatory
element. By conformational change, Ca2+-calmodulin enters into interaction with other
proteins and modulates their properties. Using this mechanism, Ca 2+ ions regulate the activity
of enzymes, ion pumps, and components of the cytoskeleton.

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