Insulin is a polypeptide hormone produced by the b-cells of islets of Langerhans of pancreas.

It has profound influence on the metabolism of carbohydrate, fat and protein.

Insulin is considered as anabolic hormone, as it promotes the synthesis of glycogen, triacylglycerols and proteins.
This hormone has been implicated in the development of diabetes mellitus.
Insulin occupies a special place in the history of biochemistry as well as medicine. Insulin was the first hormone to be
isolated, purified and synthesized; first hormone to be sequenced; first hormone to be produced by recombinant DNA
technology.

Structure of insulin
Human insulin (mol. wt. 5,734) contains 51 amino acids, arranged in two polypeptide chains. The chain A has 21 amino
acids while B has 30 amino acids. Both are held together by two interchain disulfide bridges, connecting A7 to B7 and A20
to B19. In addition, there is an intrachain disulfide link in chain A between the amino acids 6 and 11.
 +
synthesized; first hormone to be sequenced; first H 3N
hormone to be produced by recombinant DNA
technology.
Biosynthesis of insulin
Structure of insulin
Insulin is produced by the b-cells of the islets of Langerhans of
Human insulin (mol. wt. 5,734) contains 51
pancreas. amino acids, arranged in two polypeptide COO
–

chains. The chain A has 21 amino acids while B

The gene for this protein synthesis is located
has 30onamino
chromosome
acids. Both are 11.held together by
The synthesis of insulin involves two precursors, namely
two interchain disulfide bridges, connecting A7
to B7 and A20 to B19. In addition, there is an
preproinsulin with 108 amino acids (mol. intrachain
wt. 11,500) disulfideand
link in chain A between the
Preproinsulin

proinsulin with 86 amino acids (mol. wt. 9,000).

amino acids 6 and 11.
Endoplasmic
reticulum
They are sequentially degraded to form the active hormone
Biosynthesis of insulin
Insulin is produced by the -cells of the islets Signal
insulin and a connecting peptide (C-peptide).
of Langerhans of pancreas. The gene for this
sequence

Insulin and C-peptide are produced in equimolar concentration.

protein synthesis is located on chromosome 11.
The synthesis of insulin involves two precursors,
S
namely preproinsulin with 108 amino acids S S
C-peptide has no biological activity, however its estimation in the
(mol. wt. 11,500) and proinsulin with 86 amino
S S
S

acids (mol. wt. 9,000). They are sequentially

plasma serves as a useful index for the endogenous production of
degraded (Fig.36.1) to form the active hormone
insulin. insulin and a connecting peptide (C-peptide).
Insulin and C-peptide are produced in equimolar
In the b-cells, insulin (and also proinsulin) combines with zinc to
concentration. C-peptide has no biological
activity, however its estimation in the plasma Proinsulin
form complexes. In this form, insulin is stored in the granules of
serves as a useful index for the endogenous Golgi
the cytosol which is released in response to various stimuli
production of insulin. apparatus

(discussed below) by exocytosis. In the -cells, insulin (and also proinsulin)

combines with zinc to form complexes. In this 7 S S
6
7
S
11 S
form, insulin is stored in the granules of the 19 S S 20
cytosol which is released in response to various 21
stimuli (discussed below) by exocytosis. 30 A-chain
B-chain
Regulation of insulin secretion Insulin

About 40-50 units of insulin is secreted daily C-peptide

by human pancreas. The normal insulin concen-
Fig. 36.1 : Formation of insulin from preproinsulin.
tration in plasma is 20-30 U/ml. The important
• Regulation of insulin secretion
About 40-50 units of insulin is secreted daily by human pancreas. The normal insulin concentration
in plasma is 20-30 µU/ml. The important factors that influence the release of insulin from the b-
cells of pancreas are discussed hereunder.

Factors stimulating insulin secretion :

• These include glucose, amino acids and gastrointestinal hormones.
• Glucose is the most important stimulus for insulin release. The effect is more predominant when
glucose is administered orally (either direct or through a carbohydrate-rich meal). A rise in blood
glucose level is a signal for insulin secretion.
• Amino acids induce the secretion of insulin. This is particularly observed after the ingestion of
protein-rich meal that causes transient rise in plasma amino acid concentration. Among the
amino acids, arginine and leucine are potent stimulators of insulin release.
• Gastrointestinal hormones (secretin, gastrin, pancreozymin) enhance the secretion of insulin. The
GIT hormones are released after the ingestion of food.

Factors inhibiting insulin secretion : Epinephrine is the most potent inhibitor of insulin release. In
emergency situations like stress, extreme exercise and trauma, the nervous system stimulates
adrenal medulla to release epinephrine. Epinephrine suppresses insulin release and promotes
energy metabolism by mobilizing energy yielding compounds-glucose from liver and fatty acids from
adipose tissue.
Degradation of insulin
In the plasma, insulin has a normal half-life of 4-5 minutes. This short half-life permits rapid metabolic changes in
accordance to the alterations in the circulating levels of insulin. This is advantageous for the therapeutic purposes.
A protease enzyme, namely insulinase (mainly found in liver and kidney), degrades insulin.

Metabolic effects of insulin

Insulin plays a key role in the regulation of carbohydrate, lipid and protein metabolisms (Table 36.1). Insulin exerts
anabolic and anticatabolic influences on the body metabolism.
1. Effects on carbohydrate metabolism : In a normal individual, about half of the ingested glucose is utilized to meet the
energy demands of the body (mainly through glycolysis). The other half is converted to fat (~40%) and glycogen (~ 10%).
This relation is severely impaired in insulin deficiency. Insulin influences glucose metabolism in many ways. The net effect
is that insulin lowers blood glucose level (hypoglycemic effect) by promoting its utilization and storage and by inhibiting
its production.
Effect on glucose uptake by tissues : Insulin is required for the uptake of glucose by muscle (skeletal, cardiac and
smooth), adipose tissue, leukocytes and mammary glands. Surprisingly, about 80% of glucose uptake in the body is
not dependent on insulin. Tissues into which glucose can freely enter include brain, kidney, erythrocytes, retina, nerve,
blood vessels and intestinal mucosa. As regards liver, glucose entry into hepatocytes does not require insulin. However,
insulin stimulates glucose utilization in liver and, thus, indirectly promotes its uptake.
Effect on glucose utilization : Insulin increases glycolysis in muscle and liver. The activation as well as the quantities of
certain key enzymes of glycolysis, namely glucokinase (not hexokinase) phosphofructokinase and pyruvate kinase are
increased by insulin. Glycogen production is enhanced by insulin by increasing the activity of glycogen synthetase.
• Effect on glucose production : Insulin decreases gluconeogenesis by suppressing the enzymes
pyruvate carboxylase, phosphoenol pyruvate carboxykinase and glucose 6- phosphatase.
Insulin also inhibits glycogenolysis by inactivating the enzyme glycogen phosphorylase.
• 2. Effects on lipid metabolism : The net effect of insulin on lipid metabolism is to reduce the
release of fatty acids from the stored fat and decrease the production of ketone bodies.
Among the tissues, adipose tissue is the most sensitive to the action of i
• Effect on lipogenesis : Insulin favours the synthesis of triacylglycerols from glucose by
providing more glycerol 3-phosphate (from glycolysis) and NADPH (from HMP shunt). Insulin
increases the activity of acetyl CoA carboxylase, a key enzyme in fatty acid synthesis.
• Effect on lipolysis : Insulin decreases the activity of hormone-sensitive lipase and thus
reduces the release of fatty acids from stored fat in adipose tissue. The mobilization of fatty
acids from liver is also decreased by insulin. In this way, insulin keeps the circulating free fatty
acids under a constant check.
• Effect on ketogenesis : Insulin reduces ketogenesis by decreasing the activity of HMG CoA
synthetase. Further, insulin promotes the utilization of acetyl CoA for oxidation (Krebs cycle)
and lipogenesis. Therefore, the availability of acetyl CoA for ketogenesis, in the normal
circumstances, is very low.
• 3. Effects on protein metabolism : Insulin is an anabolic hormone. It stimulates the entry of
amino acids into the cells, enhances protein synthesis and reduces protein degradation.
• Mechanism of action of insulin
• It is now recognized that insulin binds to specific plasma membrane receptors present on the target tissues, such
as muscle and adipose. This results in a series of reactions ultimately leading to the biological action. Three distinct
mechanisms of insulin action are known. One concerned with the induction of transmembrane signals (signal
transduction), second with the glucose transport across the membrane and the third with induction of enzyme
synthesis.
1. Insulin receptor mediated signal transduction
• Insulin receptor : It is a tetramer consisting of 4 subunits of two types and is designated as a2b2. The subunits are in
the glycosylated form. They are held together by disulfide linkages. The a-subunit (mol. wt. 135,000) is extracellular
and it contains insulin binding site. The b-subunit (mol. wt. 95,000) is a transmembrane protein which is activated
by insulin. The cytoplasmic domain of b -subunit has tyrosine kinase activity. The insulin receptor is synthesized as a
single polypeptide and cleaved to a and b subunits which are then assembled. The insulin receptor has a half-life of
6-12 hours. There are about 20,000 receptors per cell in mammals.
• Signal transduction : As the hormone insulin binds to the receptor, a conformational change is induced in the a-
subunits of insulin receptor. This results in the generation of signals which are transduced to b -subunits. The net
effect is that insulin binding activates tyrosine kinase activity of intracellular b -subunit of insulin receptor. This
causes the autophosphorylation of tyrosine residues on b -subunit. It is believed that receptor tyrosine kinase also
phosphorylates insulin receptor substrate (IRS). The phosphorylated IRS, in turn, promotes activation of other
protein kinases and phosphatases, finally leading to biological action .
• Insulin-mediated glucose transport : The binding of insulin to insulin receptors signals the translocation of vesicles
containing glucose transporters from intracellular pool to the plasma membrane. The vesicles fuse with the
membrane recruiting the glucose transporters. The glucose transporters are responsible for the insulin–mediated
uptake of glucose by the cells. As the insulin level falls, the glucose transporters move away from the membrane to
the intracellular pool for storage and recycle.
• Insulin mediated enzyme synthesis : Insulin promotes the synthesis of enzymes such as glucokinase,
phosphofructokinase and pyruvate kinase. This is brought about by increased transcription (mRNA synthesis),
followed by translation (protein synthesis).
 transduction (IRS—Insulin receptor substrate).Insulin promotes the synthesis of enzymes such
activity of intracellular -subunit of insulinphosphofructokinase and
as glucokinase,
pyruvate kinase. of
receptor. This causes the autophosphorylation This is brought about by
are transduced to -subunits. The net effect is increased transcription (mRNA synthesis),
tyrosine
that insulin bindingresidues
activates on -subunit.
tyrosine kinase Itfollowed
is believed that (protein synthesis).
by translation
Membrane receptor tyrosine kinase also phosphorylates
insulin receptor substrate (IRS). The phospho-
Tyr Tyr rylated IRS, in turn, promotes activation
Glucose of other
Cytoplasm () () ()
Insulin protein kinases and phosphatases, finally leading
( )
to biological action (Fig.36.2Glucose
).

()

()
2.Insulin
Insulin-mediated glucose transport : The

()
()
()

()
binding of insulinSignal to insulin receptors signals the
Translocation
translocationInsulin- of vesicles containing glucose
()
Fission
Membrane receptor
transporters complexfrom intracellular pool to the Plasma
plasma membrane. The vesicles fuse with the membrane
() ()
IRS—Tyr

()
()
Tyr Tyr
Cytoplasm
IRS—Tyr membrane recruiting the glucose transporters.
The glucose transporters are responsible for the

()

()
P P

()

()
P insulin–mediated uptake of glucose by the cells.
Glucose transporters
As the insulin level falls, the glucose transporters
move away from the pool
Intracellular membrane
of vesicles to the
Biological Protein kinases
effects Phosphatases intracellular pool Fig. 36.3for storage
: Insulin and transport.
mediated glucose recycle
(activated) (Fig.36.3).

Fig. 36.2 : Insulin receptor mediated signal 3. Insulin mediated enzyme synthesis :
transduction (IRS—Insulin receptor substrate). Insulin promotes the synthesis of enzymes such
as glucokinase, phosphofructokinase and
 Glucagon

• Glucagon, secreted by a-cells of the pancreas, opposes the actions of insulin.

• It is a polypeptide hormone composed of 29 amino acids (mol. wt. 3,500) in a single chain.
• Glucagon is actually synthesized as proglucagon (mol. wt. 9,000) which on sequential degradation releases active
glucagon. Unlike insulin, the amino acid sequence of glucagon is the same in all mammalian species (so far studied).
Glucagon has a short half-life in plasma i.e. about 5 minutes.
• Regulation of glucagon secretion
The secretion of glucagon is stimulated by low blood glucose concentration, amino acids derived from dietary
protein and low levels of epinephrine. Increased blood glucose level markedly inhibits glucagon secretion.
• Metabolic effects of glucagon
Glucagon influences carbohydrate, lipid and protein metabolisms. In general, the effects of this hormone oppose
that of insulin.

• Effects on carbohydrate metabolism : Glucagon is the most potent hormone that enhances the blood glucose level
(hyperglycemic). Primarily, glucagon acts on liver to cause increased synthesis of glucose (gluconeogenesis) and enhanced
degradation of glycogen (glycogenolysis). The actions of glucagon are mediated through cyclic AMP.
• Effects on lipid metabolism : Glucagon promotes fatty acid oxidation resulting in energy production and ketone body
synthesis (ketogenesis).
• Effects on protein metabolism : Glucagon increases the amino acid uptake by liver which, in turn, promotes
gluconeogenesis. Thus, glucagon lowers plasma amino acids.
Mechanism of action of glucagon
Glucagon binds to the specific receptors on the plasma membrane and acts through the mediation of cyclic
AMP, the second messenger.
Cyclic AMP (cAMP, cyclic adenosine 3’,5’-monophosphate) is a ubiquitous nucleotide.
It consists of adenine, ribose and a phosphate (linked by 3’,5’ linkage).
cAMP acts as a second messenger for a majority of polypeptide hormones.
Chapter 19 : HORMONES 431
The membrane-bound enzyme adenylate cyclase converts ATP to cyclic AMP. cAMP is hydrolysed by phosphodiesterase
to 5’-AMP ATP hormonal action. In response to the stimuli of
Adenylate central nervous system, hypothalamus liberates
cyclase certain releasing factors or hormones. These
PPi 2+
Mg factors stimulate or inhibit the release of
corresponding tropic hormones from the anterior
NH2
pituitary. Tropic hormones stimulate the target
N endocrine tissues to secrete the hormones they
N
synthesize. The relationship between
N N hypothalamus and pituitary with endocrine
glands is illustrated in Fig.19.6. In general, the
5 hormonal system is under feedback control. For
O CH2 O
instance, adrenocorticotropic hormone (ACTH)
– H H inhibits the release of corticotropin releasing
O P O H H
3 hormone (CRH).
O OH
3 ,5 -Cyclic adenosine
monophosphate (cAMP) HYPOTHALAMIC HORMONES
H2O
Phosphodiesterase
Hypothalamus produces at least six releasing
5 -AMP
factors or hormones.
 general view response

Once system
Adenylate cyclase produced, cAMP
A series of performs itsatrole
events occur as a second
the membrane Fig. 19.2the
level that influence : Mechanism
activity of adenylate
of action ofcyclase leading (H–Horm
steroid hormones
messenger
to the synthesis of cAMP. in elicitingis mediated
This process bio- by G-proteins, R–Receptor; HR–Hormone-receptor
so designated due to their abilitycomplex).
to
bind to guanine responses (Fig.19.4).
nucleotides.
chemical
cAMP activates protein kinase A
Action of (AcAMP—a
stands general
for cAMP).viewThis enzyme is a heterote- protein that ultimately causes the bioch
tramer
Once produced, cAMPconsisting of role
performs its 2 regulatory
as a secondsubunits
messenger(R) response. responses.
and biochemical
in eliciting
2 catalytic
cAMP activates subunits
protein kinase (C). for cAMP).
A (A stands
It should,
This enzyme is a heterotetramer consisting of 2 regulatory subunits (R) and 2 catalytic subunitshowever,
(C). be remembere
cAMP binds to inactive protein kinase and cAMP does not act on all protein kinas
cAMP binds causes the dissociation
to inactive protein kinaseofandR causes
and C the subunits.
dissociation of instance, on protein kinase C (the
R and C subunits.
messenger is diacylglycerol).
4cAMP + R2C2 R2(4 cAMP) + 2C
(inactive) (inactive) (active) Dephosphorylation of proteins : A g
The active subunit
The (C) catalyses
active phosphorylation
subunit (C) catalyses of proteins
phosphory- (transferenzymes called
of phosphate groupprotein phosphatases
to serine and hy
threonine residues).
lation of proteins (transfer of phosphate group to and remove the phosphate group ad
It is the phosphoprotein that ultimately
serine and threonine causesIt the
residues). biochemical
is the phospho-response.proteins.
It should, however, be remembered that cAMP does not act on all protein kinases. For instance, on protein
kinase C (the second messenger is diacylglycerol).
Dephosphorylation of proteins : A group of enzymes called protein phosphatases hydrolyse and remove the
phosphate group added to proteins
Degradation of cAMP : cAMP undergoes rapid hydrolysis, catalysed by the enzyme phosphodiesterase to 5’ AMP
which is inactive. Hence, the effect of cAMP will be shortlived if the hormone stimulating adenylate cyclase is
removed. Caffeine and theophylline (methylxanthine derivatives) can inhibit phosphodiesterases and increase the
intracellular levels of cAMP.432 BIOCHEMISTRY

Hormone

Receptor

GTP regulatory protein Plasma membrane

Adenylate
cyclase
Cytosol
ATP PPi

Phosphodiesterase R C
AMP 4cAMP
( )
R C

R2 C2

R
C C
R

R 2 (4 cAMP) ATP ADP

Protein Phosphoprotein

Phosphatase

Ultimate
Pi biochemical
response