BIOCHEMISTRY

HORMONAL REGULATION II
Dr. Suzette M. Mendoza | March 05, 2019 LE5 TRANS3

OUTLINE  Insulin decreases blood glucose levels by stimulating glucose

I. Overview II. Clinical Correlations uptake into muscle, adipose tissue, and liver
II. Insulin A. Overview of Insulin  Not all organs need insulin for glucose to enter the cells
A. Structure Deficiency/Resistance  These tissues have GLUT 1, GLUT 2, GLUT 3, and GLUT 5
B. Synthesis and B. Type I Diabetes (located at the plasma membrane)
Degradation Mellitus  Examples:
C. Regulators of C. Type II Diabetes  Brain – due to GLUT 1 transporter
Insulin Release Mellitus  Retina
D. Release of Insulin D. Monogenic forms of  Renal Medulla
E. Action of Insulin Diabetes Mellitus  Liver: has GLUT 2, therefore not an insulin-dependent organ for
F. Degradation of E. Gestational Diabetes glucose entry
Insulin Mellitus  The function of insulin in the liver is to regulate glycolysis by
G. Signaling Cascade F. Neonatal Diabetes upregulating active glucokinase
of Insulin Mellitus  Glucokinase is initially inactive in the nucleus
H. Insulin in III. Glucagon o Upon increase of blood glucose and insulin in the
Carbohydrate A. Action of Glucagon liver, glucokinase translocation occurs from the
Metabolism B. Synthesis of cytosol to the nucleus
I. Insulin in Lipid Glucagon
Metabolism C. Structure of Glucagon Insulin at Target Tissue
J. Insulin in Protein D. Glucagon Receptor  Insulin-dependent organs utilize GLUT 4 for glucose entry while
Metabolism E. Degradation of simultaneously propagating a variety of intracellular responses
K. Regulation of Glucagon
 In the absence of insulin, GLUT 4 is located in
Insulin Signaling F. Regulation of
intracytoplasmic vesicles (unlike other GLUTs which are
Cascade Glucagon
already on the plasma membrane constitutively)
L. Termination of G. Release of Glucagon
Signaling Pathway H. Signaling Cascade of
 Insulin-Dependent Organs
of Insulin by Glucagon
Inhibitors I. Downregulation of  adipose tissues
Glucagon  skeletal muscles
IV. Summary  cardiac muscles
V. Incretins
VI. References  Receptor Tyrosine Kinase
VII. Appendix 1. Insulin binds to Receptor Tyrosine Kinase (RTK)
a. RTK is composed of 2 extracellular  subunits and 2
transmembrane  subunits
OBJECTIVES i. Quaternary structure maintained by disulfide
 Review the metabolic effects of Insulin and Glucagon bonds
 Expound the biochemistry of Insulin and Glucagon b. RTK has intrinsic phosphorylation activity to activate
 Correlate the signaling pathways of Insulin and Glucagon with signaling cascade
their metabolic effects i. Cross phosphorylation of  subunit activates
 Analyze the biochemical bases of different diseases due to signaling cascade when insulin binds to the 
Insulin deficiency or resistance subunit of the insulin
 Describe the factors that can terminate the signaling pathway of
Insulin and Glucagon  Insulin activated secondary messengers and response
 Discuss the functions of Incretins to regulate metabolism  Activate AKT
I. OVERVIEW  GLUT 4 fusion with plasma membrane
 Active AKT inhibits GSK-3
 Insulin and glucose activities are primarily dependent on blood o Inactive GSK-3 stimulates Glycogen Synthesis
glucose levels  Upregulate SREBP
 Glucose homeostasis  Enhance cholesterol and FA synthesis
 Normal blood glucose: 4.5mM = 72-126 mg/dL  Activate Sik2
 actions of major hormones regulating fuel metabolism  Inhibit Gluconeogenesis
(insulin, glucagon, epinephrine and cortisol) on metabolic  Activate PDE3B
processes in many body tissues
 Inhibit lipolysis
 Total glucose requirements/day:
 Activate mTORC1 (atypical serine/threonine kinase)
 Brain – 150 g
 Cell Survival
 Tissues – 40 g
 Metabolism
 Major target organs of insulin:
 Liver  Proliferation
 Skeletal Muscles  Activate Ras pathway
 Adipose tissues  Growth
 Organs that only use glucose for energy: Table 1. Major hormones that regulate fuel metabolism
 Brain ANABOLIC CATABOLIC
 Mature RBC Insulin Glucagon
 Kidney Medulla Epinephrine and norepinephrine
 Lens Glucocorticoids
 Testis
L.E. # 5 - Trans # 3 Group Q: Tagum, Tan, B., Tan, D., Tan,L., Tan, R. 1 of 11
 Thyroid hormones
Growth hormones
Somatostatin
Biochemistry of Insulin and Glucagon
 Class II hormones
 Peptide hormones hydrophilic
 Can transport freely through blood, no carrier protein needed
 Polypeptide hormones are synthesized as large hormones
initially. Preprohormones undergo a series of proteolysis to
obtain the mature form.
Cell secretion
 Insulin secreted from  cells Islet of Langerhans
 Glucagon secreted from  cells in Islets of Langerhans
Type of Receptor
 Insulin: Receptor Tyrosine Kinases
 Glucagon: GPCR
II. INSULIN
 Major anabolic hormone
 Insulin signaling system coordinates systemic growth
 Released from the β-cells of the pancreas in response to
carbohydrate ingestion, promotes glucose utilization as a fuel
and glucose storage as fat and glycogen[2021A trans]
 At molecular levels insulin regulates many pathways
 Glycolysis and glucose storage (muscle and liver)
 Inhibits ketogenesis and gluconeogenesis (liver)
 Lipid synthesis and storage (liver)
 Stimulation of protein synthesis (muscle and liver)
 Insulin increases as blood glucose level rises
 ~30-45 minutes after a high carbohydrate meal intake (well-
fed nutritional stage)
 Insulin decreases as blood glucose level diminishes
 Glucose is absorbed and stored or utilized by tissues
 Blood glucose return to basal level (~120 minutes after
meal)
 Metabolic effects:
 Carbohydrate Metabolism
 Increasing glucose uptake in skeletal muscles & adipocytes by
enhancing GLUT4 on the PM
 Decrease Gluconeogenesis
 Lipid Metabolism
 Increased lipid synthesis in liver & adipocytes (via SREBP)
 Decreasing FA release from TAGs in adipocytes & muscles
 Protein Metabolism
 Increase protein synthesis
 Decrease protein degradation
A. STRUCTURE OF INSULIN
Figure 1. Structure of human insulin
Biochemistry Hormonal Regulation of Metabolism 2
 Polypeptide hormone with 2 chains of 51 AAs
 Quaternary structure
 A chain (21 AAs) : B chain (30 AAs)
 Linked by 3 disulfide bridges (S-S bridges) contributed by
cysteine residues
 2 inter-chain S-S bridges (connects A7 to B7 & connects
A20 to B19)
 1 intra-chain S-S bond (connects AAs 6 to 11 of the A chain)
B. SYNTHESIS AND DEGRADATION OF INSULIN
Synthesis of Insulin
Figure 2. a) Insulin synthesis b) mature insulin
 Preproinsulin
 MW: 11,500 kD
 Pre or leader sequence
 23 hydrophobic amino acids (core hydrophobic amino
acids) that direct the molecule into the cisternae of ER
 Proinsulin
 MW: 9,000 kD
 Provides the conformation for the proper location of the
disulfide (S-S) bridges
 Sequence: B chain – C (connecting) peptide – A chain
 Mature Insulin
 Composed of an A chain and a B chain linked by 3 disulfide
bridges
 C-peptide is representative of mature insulin since they are
secreted together
 C-peptide can be used in tests to measure insulin since
insulin has a very short half-life
Modification:
Signaling sequence cleaved in ER
C-Peptide cleaved in Golgi apparatus
Degradation of Insulin
 Since insulin is a polypeptide hormone, it can be degraded by
denaturation or cleavage of peptide bonds
 Enzyme: insulinase
 Site: primarily in the liver and to a lesser extent in the kidneys and
skeletal muscles
C. REGULATORS OF INSULIN RELEASE
 Glucose levels in the pancreas stimulate insulin release.
 Increased glucose levels or hyperglycemia are the major
stimulating factors of insulin release
 Only insulin & glucagon are synthesized and released in direct
response to changing levels of sugar in the blood.
 Threshold for insulin release: 80 mg%
 Proportional to glucose concentration up to 300 mg%
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Table 2. Major and Minor Regulators of Insulin Release Second Phase
REGULATORS EFFECT  Happens hours after eating because insulin must be
MAJOR REGULATORS synthesized and released
Glucose +  Increased cytosolic acetyl CoA concentration triggers release of
MINOR REGULATORS newly-synthesized insulin from the β-cells
Amino Acids +
Neural input +
GUT hormones (incretins, GIP, +
GLP-1)
Epinephrine -

D. RELEASE OF INSULIN

Figure 5. Phases of Insulin release

E. ACTION OF INSULIN

Insulin Receptor (IR)

 Receptor tyrosine kinases (RTKs)
 IGF [Insulin-like Growth Factor] and IRR [Insulin Receptor-
Related Receptor]
 Proteolytic cleavage of a single polypeptide chain precursor
 α & β chains linked by disulfide bonds
 Biologically active α2β2 receptor heterotetramer
 Preformed heterotetramer:
 2 extra-cellular α subunits: insulin binding domains
 Inhibits the tyrosine kinase activity of the β subunit
 2 transmembrane β subunits:
 Joined by S-S bridges
 Tyrosine Kinase activity and autophosphorylation sites
on the β subunits
Figure 3. Steps in insulin release  With 2 domains:
 Transmembrane domain
Mechanism of insulin release:  Intracytoplasmic domain (contains the ATP binding site
1. Glucose is transported into the pancreatic β-cell through and where RTK property resides)
GLUT 2
2. Glucose is oxidized yielding ATP
3. Increase in ATP/ADP ratio inhibits ATP gated K+
Channels, causing the cell to depolarize
4. Depolarization opens VG calcium channels
5. Increase in calcium causes exocytosis of insulin from
vesicles

First Phase
 Insulin is released from the pancreas in a biphasic manner
 First phase is a brief spike (immediately after eating) in
insulin followed by a post prandial plateau (hours)
 Immediate response to increased blood glucose levels
 First phase response involves release of pre-formed insulin
ATP-Gated K+ Channels
 Molecular target of sulfonylurea (anti-diabetic drug)
 Closes when sulfonylurea is present and causes insulin
release

Figure 6. Insulin Receptor

 Only 3 are expressed in humans
 IRS1 and IRS2
 Expressed in the brain, muscle, heart, adipocyte, liver
kidney, ovary and mammary gland
 Physiologic functions include:
Figure 4. Structure of KATP Channels  Pancreatic β cell growth and function
Biochemistry Hormonal Regulation of Metabolism 2 3 of 11
  Central nutrient sensing F. DEGRADATION OF INSULIN
 Cancer progression and lifespan  Half-life: 3-5 minutes
 IRS4  Occurs in the liver and to a lesser extent in the kidneys and also
 Largely restricted to the hypothalamus and thymus the placenta
 50% of the degradation takes place in the liver
Metabolic Effects of Insulin  Involves two enzymes:
 Diverse biologic responses by binding of insulin to IR  Protease (Insulinase): Specific enzyme
 Glucose transport into tissue, decrease blood glucose levels  Glutathione-insulin transhydrogenase: destabilizes
 Glycogen synthesis disulfide bonds
 Lipid synthesis  Endocytosis of IR complex
 Protein synthesis G. SIGNALING CASCADE OF INSULIN
 Mitogenesis
 Upon the binding of insulin, protein tyrosine kinase activity of the
 Cell survival
receptor is enhanced, inducing phosphorylation of diverse target
proteins.
Insulin Enzymatic Effects
 The adaptor protein IRS1 is a major substrate of the insulin
 Key regulatory enzymes are controlled via covalent modification
receptor, serving as a platform for the signaling complex.
2 mechanisms:
 Phosphoinositide 3-kinase (PI3K) is a component of this
 Activation of protein phosphatases
signaling complex, catalyzing phosphorylation in position 3 of the
 Decrease intracellular cAMP inositol ring of phosphoinositides.
 inactive PKA (regulatory subunit) because cAMP activates  Products of PI3K, such as PIP3 and PIP2, bind to pleckstrin
PKA homology domains (PH) kinases, such as PDK1 and Akt2 or
Protein Kinase B (PKB), recruiting them to the plasma
Insulin effect on enzymes membrane.
 Activated enzymes:  A specific serine residue of the carboxyl-terminal hydrophobic
 PFK1, pyruvate kinase, PFK2: enzymes associated w/ motif of Akt2 is phosphorylated
glycolysis  then translocated PDK1 phosphorylates threonine in the
 Glycogen synthase: key regulatory enzyme of glycogenesis activation segment of Akt2, leading to the full activation of
 PDH: only enzyme of the PDH reaction Akt2
 Acetyl CoA carboxylase: key enzyme for fatty acid synthesis
Insulin Signaling Pathway
 Inactivated enzymes:
 Glycogen phosphohrylase, phosphorylase kinase:  Insulin receptor is active after phosphorylation of critical tyrosine
enzymes responsible for glycogenolysis residues (this is the start of any cascade)
 Fructose 2,6-bisphosphatase: key enzyme of  Activated tyrosine receptors will phosphorylate insulin receptor
gluconeogenesis substrates (IRS) 1 to 4 (more important are 1 and 2; IRS 4 is
found in the hypothalamus and thymus)
 Hormone sensitive lipase: enzyme in charge of lipolysis
 IRS must have phosphotyrosine domains which are the ones
to be phosphorylated to give activated IRS
Induction/Repression  IRS: immediate downstream substrate of a
 Insulin can induce or repress translation of key enzymes, phosphorylated/active insulin receptor
therefore affecting enzyme concentration  Inhibitors of phosphorylation of IRS:
 Phosphotyrosine phosphatase (PTP)
Table 3. Insulin-induced enzymes  SHP-2
INDUCTION OF KEY ENZYMES  SRC homologue phosphatase
ENZYME METABOLIC EFFECT  Type-1 diabetes is characterized by the inability to synthesize
Glucokinase ↑ Glycolysis insulin
Hexokinase  Type-2 diabetes the body becomes resistant to the effects of
PFK1 insulin
PFK2
Pyruvate kinase H. INSULIN IN CARBOHYDRATE METABOLISM
Glucose-6-dehydrogenase ↑ NADPH GLUT4 Translocation
6-Phosphogluconate Stimulate FA and  PI3K-Akt Dependent Pathway (skeletal muscle and adipose
dehydrogenase Cholesterol synthesis tissue)
Malic enzyme 1. Insulin binds to the α-subunit of the RTK-IR
Acetyl CoA carboxylase ↑ Fatty Acid synthesis 2. RTK β subunit will be phosphorylated
9 Desaturase 3. RTK β subunit binds and phosphorylates IRS1
3-HMG CoA reductase ↑ Cholesterol synthesis 4. IRS1 phosphorylates and activates Gab1 (adaptor protein
Glycerol-3-phosphate ↑ TAG & phospholipid with HS domains) and p85 (regulatory subunit of PI3K)
acyltransferase synthesis  p110, the catalytic subunit of PI3K, is now active
Lipoprotein lipase TAG (CM & VLDL)  Gab1-p85: docking site for p110
FFA + Glycerol 5. PI3K phosphorylates PIP2, a plasma membrane lipid, to
Table 4. Insulin-repressed enzymes
form PIP3
REPRESSION OF KEY ENZYMES 6. PIP3 phosphorylates PDK1
ENZYME METABOLIC EFFECT 7. PDK1 phosphorylates Akt2 and atypical PKCλ/ζ
PEPCK Inhibits Gluconeogenesis  PDK1 and Akt2 are serine-threonine kinases
Pyruvate dehydrogenase 8. Akt2 phosphorylates several anchoring proteins (ie. AS160
kinase coupled to an anchoring protein 14-3-3 and TBC1D1)
Pyruvate carboxylase  AS160 and TBC1D1 are GTPases
Glucose-6-phosphatase  GLUT4 in the cytoplasm are bound to GDP
Aminotransferases  Complex will act on GLUT4 to replace GDP with GTP

Biochemistry Hormonal Regulation of Metabolism 2 4 of 11

 9. Conformation will translocate GLUT4 from cytosol to
plasma membrane
 PI3K-Akt Independent Pathway (adipose tissue)
1. Active IRS phosphorylates Cbl, a ubiquination factor
recruited by anchoring protein CAP, forming the Cbl-CAP
complex
 Phosphorylation of Cbl requires another protein that
recruits Cbl to the insulin receptor, the adaptor protein APS
containing PH and SH2 domains
2. Cbl-CAP-APS complex translocates in lipid rafts in the
plasma membrane
 Flotillin (hydrophobic lipid raft protein) associates with CAP
and then CAP is localized in the lipid rafts
3. Cap-Cbl-APS complex recruits cRK II
4. cRK II activates C3G, a nucleotide exchange protein
5. C3G facilitates the replacement of GDP with GTP, which
will activate the members of the GTP-binding protein family,
TC10
6. TC10 promotes the translocation of GLUT4 to the plasma
membrane
Stimulation of Glycogenesis
1. Phosphorylated and active Akt enters cytoplasm from
plasma membrane
2. Akt phosphorylates and inactivates glycogen synthase
kinase 3 (GSK3)
 A major substrate of GSK3 is glycogen synthase (GS), an
enzyme that catalyzes the final step in glycogen
synthesis
 Phosphorylation of glycogen synthase by GSK3 inhibits
glycogen synthesis
Inactivation of GSK3 by AKT via phosphorylation, promotes
glucose storage as glycogen (glycogenesis)
Inhibition of Gluconeogenesis
 Opposing effects of glucagon and insulin on the cAMP-
responsive element (CRE)-binding protein (CREB) pathway.
1. Glucagon stimulates the protein kinase A (PKA)-mediated
phosphorylation of CREB at Ser133.
2. PKA stimulates cAMP-regulated transcriptional co-
activator 2 (CRTC2) activity via the phosphorylation and
inhibition of salt-inducible kinase 2 (SIK2), leading to the
dephosphorylation of CRTC2.
3. Dephosphorylated CRTC2 translocates to the nucleus,
where it binds to CREB and promotes the recruitment of
TBP-associated factor 4 (TAF4) and CREB-binding protein
(CBP) and its paralogue p300.
4. Nuclear CRTC2 is transiently stabilized by CBP/p300-
mediated acetylation (Ac) at Lys628.
5. Insulin signaling stimulates a member of the AKT family,
which phosphorylates and activates SIK2. Active SIK2
disrupts the CRTC2–CBP/p300 interaction by
phosphorylating CBP/p300 at Ser89, leading to the
deacetylation and ubiquitin (Ub)-dependent degradation of
CRTC2. SIK2 also promotes the cytoplasmic translocation
of CRTC2 through phosphorylation at Ser171.
I. INSULIN IN LIPID METABOLISM
 Lipid and protein synthesis: starts with mTORC1 (with variance
in the following step)
 Lipid synthesis: uses SREBP-1
 PI3K and Akt: required for gluconeogenesis and lipogenesis
 mTORC1: required for lipogenesis (via induction of SREBP-1c
activity)
 Cleaved SREBP1: transcription factor, promotes the expression
of diverse genes with important roles in lipid synthesis
 Examples:
 FASN (fatty acid synthase)
 GPAT (glycerol-3-phosphate acyltransferases)
 ACLY (ATP citrate lyase)
 ACC (acetyl-CoA carboxylase)
Biochemistry Hormonal Regulation of Metabolism 2
 SCD1 (stearoyl-CoA desaturase 1)
 GK (glucokinase)
Stimulation of Lipogenesis
1. mTORC2 stimulates Akt2
2. Akt2 (with PDK1 and PKCλ/ζ) stimulates transcription factor
SREBP1
3. Activated SREBP1 is translocated into the nucleus (signals
lipid synthesis)
Inhibition of Lipolysis
1. Akt2 phosphorylates and activates PDE3B
2. PDE3B degrades cAMP to AMP → inhibition of the
stimulation of cAMP-dependent PKA
3. Inactive PKA → no HSL activation → inhibition of lipolysis
J. INSULIN IN PROTEIN METABOLISM
 Lipid and protein synthesis: starts with mTORC1 (with variance
in the following step)
 Protein synthesis: uses p70 S6K
 Insulin: activates Akt/PKB and mTORC leading to protein
synthesis and cell growth
 TSC
 Heterodimer made up of TSC1 (hamartin) and TSC2 (tuberin)
 Important sensor in the regulation of mTORC1
Stimulation of Protein Synthesis
 mTORC1 Pathway
1. Activated Akt2 → inhibits dimerization between TSC2 and
TSC1
 No dimerization between the two → no inactivation of
Rheb → (+) effect: activated Rheb & retained GTP
attached to Rheb
2. Activated Rheb stimulates mTORC1
3. Under nutrient abundance, mTORC1 phosphorylates and
activates p70 S6K while phosphorylating 4EBP1 (two
simultaneous events)
4. Phosphorylation of 4EBP1 inhibits it from binding to eIF4E
(eukaryotic Initiation Factor 4E)
5. Stimulation of p70 S6K and inhibition of 4EBP1 à protein
synthesis
 mTORC2 Pathway
1. Activated mTORC2 activates Akt2
2. Akt2 phosphorylates and inactivates Proline-Rich Akt
Substrate 40 (PRAS40)
3. Inactivation of PRAS40 → reduction of interaction with
mTORC1 → activation of mTORC1 signaling
Ras and MAP Kinase Pathway
 Regulation of proliferation and differentiation of several cell
types
 Regulation of gene transcription by MAP kinase cascade
 Steps involved:
 Ligand binds to RTK which catalyzed autophosphorylation
 GRB2 then binds to phosphorylated RTK
 mSOS binds to GRB2 and membrane bound protein inactive
Ras on the other side which is initially bound to GDP
 mSOS catalyzes exchange of GDP to GTP which leads to
activation of Ras
 Active Ras binds to cRaf-1 which in turn phosphorylates
MEK 1 and 2 and ERK1 and 2 via MEK/MAP kinase
 Leads to cell growth, DNA synthesis, and early response
genes
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Figure 7. Ras and MAP Kinase Pathway
K. REGULATION OF INSULIN SIGNALING CASCADE
 Serine-threonine kinases
 Phosphorylates and inhibits IRS 1-4, no phosphorylation of
activated insulin receptor
 Negative feedback mechanism on the signaling pathway via
serine phosphorylation of IRS by interfering with functional
domains of IRS
 Protein phosphatase 2A (PP2A)
 Inhibition of MAP kinase pathway
 Key negative regulator of PI3K/Akt pathway by
dephosphorylation and inactivation of Akt/PKB and PKCλ
 Phosphotyrosine phosphatase 1B (PTP1B)
 Interacts directly with IR and dephosphorylates tyrosine
residues leading to decreased activity
 Termination of insulin action
 Phosphatase and Tensin homologue on chromosome 10
(PTEN)
 3’ phosphatase that converts PIP3 to PIP2
 Suppresor of Cytokines (SOCS) and Growth factor-receptor-
bound protein 10 (Grb10)
 Down-regulate IR function by dephosphorylation of IR and its
substrates
 SOCS might induce ubiquitin-mediated degradation of IRS1
and IRS2
 SRC homology 2 (SHIP)
 Degrades PIP2 and PIP3
 SHIP1 has a 3’ phosphatase activity
 SHIP2 has a 5’ phosphatase activity
 SH2-domain containing tyrosine phosphatase 2 (SHP2)
 Dephosphorylate other phosphotyrosines that mediate
binding of PI3K and Grb2
 Insulin degrading enzyme (IDE)
 Promotes degradation of insulin
 Internalization of insulin-insulin receptor complex into
endosomes
 Termination of insulin signaling
Biochemistry Hormonal Regulation of Metabolism 2
L. TERMINATION OF SIGNALING PATHWAY OF INSULIN
BY INHIBITORS
Figure 8. Insulin Signaling Pathway Termination. Orange circles are shown to
inhibit their respective pathways.
II. CLINICAL CORRELATIONS
A. OVERVIEW OF INSULIN DEFICIENCY/RESISTANCE
 Can be categorized based on site of resistance:
 Pre-receptor
 Possible defect
 insulin receptor (IR)
 antibodies (type B insulin resistance)
 abnormal molecule
 Rare
 Receptor
 Possible defect: decrease in the number or affinity of IR
 Common is obese patients (happens in their adipocytes)
 Post-receptor
 Possible defect: defects in signal transduction (e.g.
defective tyrosine phosphate, mutations in genes coding for
IRS-1 and PI-3K, defective translocation of GLUT4,
increase in fatty acids)
 Post-receptor resistance is the most common type of
insulin resistance
B. TYPE I DIABETES MELLITUS
 Lack of insulin release due to destruction of islet β cells of the
pancreas
 Pathophysiology of Type I DM:
 The lack of insulin leads to two lines of effects
 Increase gluconeogenesis and decrease glucose uptake
 hyperglycemia  glycosuria  osmotic diuresis 
dehydration (can be fatal)
 Increase lipolysis  increase ketogenesis  ketonemia
 ketonuria and metabolic acidosis  metabolic
acidosis leads to compensatory hyperventilation
 Onset is usually at earlier years of age because the infections
that cause Type I DM usually occur early in life (e.g. viral
infections)
 See Table 5 for a tabulated comparison between Type I and II
DM.
C. TYPE II DIABETES MELLITUS
 Due to impaired β cell function and decrease in the
sensitivity of the receptors for insulin
 Insulin synthesis is active but the receptors are unresponsive
 Will initially cause hyperinsulinemia (overproduction of insulin)
to try to compensate for the lack of unresponsiveness
 However, β cells will eventually be burned out and slow down
production of insulin  increased gluconeogenesis
 There is also hyperglycemia toxicity which can cause β cell
destruction
 Patient characteristics:
 Have high BP (140/90 mmHg or greater)
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  Have a high density lipoprotein (HDL) cholesterol level ≤ 35 Obesity Uncommon Common
mg/dL and/or a triacyglycerol (TAG) level ≥ 250 mg/dL Ketoacidosis Yes Rare (can be
 Have had impaired glucose tolerance test (GTT) or impaired precipitated by
fasting glucose on previous testing major metabolic
 Pathophysiology of Type II DM: stresses, such as
infection, sepsis,
 Factors that result in an increase in blood glucose levels:
and stress)
 Carbohydrate intake (β cells will increase insulin secretion
Treatment Insulin Hypoglycemic drugs
to try to overcome high glucose levels from increase *Recent trends/tide: and insulin
carbohydrate intake, which leads to insulin resistance of oral hypoglycemic (no matter how
the receptors) and β cell many drugs are
 Decreased peripheral glucose uptake (adipocytes and transplant (mildly administered, if no
skeletal muscles) successful) lifestyle
 Decreased insulin secretion modification, it will
 Increased hepatic glucose production still be untreated)
 Risk factors for developing Type II DM:
D. MONOGENIC FORMS OF DIABETES MELLITUS
 Obesity
 Have a relative with DM  Recently researchers say that it is a polygenic disease
 Belong to a high-risk ethnic population (African-American,  Forms associated with insulin resistance
Native American, Hispanic, or Native Hawaiian)  Mutations in the insulin receptor gene
 Have been diagnosed with gestational diabetes mellitus or  Type A insulin resistance (more common than Type B)
have delivered a baby weighing > 9 lbs (4 kg)  Leprechaunism
 The increase in size of the baby is due to the abundance of  Donohue syndrome (severe form of Leprechaunism)
glucose, which allows the baby to keep on growing  Clinical manifestations:
 Type II DM has a heritability of > 50% o Organomegaly
 Genes associated with Type II DM are summarized in Table 4. o Macrogenitalia
o Macroglossia
Table 4. Genes associated with Type II DM. o Hyperglycemia (due to insulin resistance)
Genes Comments  Rabson-Mendenhall syndrome
Peroxisomal proliferator Lipid Synthesis  Milder form of leprechaunism
activated receptor γ  Clinical manifestations:
(PPARγ)  lack of fatty tissue under the skin
IRS-1 Impaired peripheral response  atrophy of muscles
to insulin  dental abnormalities
K-inwardly-rectifying Affects insulin secretion;  excessive body hair growth (hirsutism)
channel subfamily J Codes for 2 CHONs w/c  low lying hairline
member 11 (KCNJ11) constitute ATP-sensitive K+  prominent, widely spaced eyes; broad nose; and large,
channel (see neonatal DM) low-set ears.
WFS1 Codes for Wolframin, a protein  Lipoatrophic diabetes
detected in patients w/ D.I.,  In patients who are injecting insulin, there is atrophy of
juvenile DM, deafness and muscles due to insulin
optic atrophy; Usually the  Mutations in the PPARγ gene
younger the patient, the more  Forms associated with defective insulin secretion
severe the manifestation  Mutations in insulin or proinsulin genes
because insulin also  Mitochondrial gene mutations
contributes to growth and  Maturity-onset diabetes of the young (MODY)
development  Characterized by persistent connecting or C-peptide
HNF1 homeobox A Associated with Maturity Onset secretion
(HNF1A) & HNF1 Diabetes in the Young (MODY)
homeobox B (HNF1B) Nice to know:
SLC30A8 Codes for zinc transporter  HNF: Hepatocyte nuclear factor
FTO Associated with obesity  IPF: Insulin promoter factor
GCKR & IGF1 Associated w/ Insulin  NeuroD1/BETA2: neurogenic differentiation 1 / beta cell E-
resistance box trans-activator 2
 PPAR: peroxisome proliferator-activated receptor
Table 5. Summary of similarities and differences between Type I and II DM.
 HNF-4α = MODY 1
Characteristics Type I DM Type II DM
 Glucokinase = MODY 2
Onset <20 years of age >40 years of age
o Most common type of MODY
Insulin synthesis Absent: Immune Preserved: Impaired
destruction of islet β β cell function and
o No glucokinase  no production of ATP from glucose
cells (insulin insulin resistance  no closure of ATP-sensitive K+ channel (constant
deficiency) (e.g. due opening)  no insulin secretion
to infection)  HNF-1α = MODY 3
Plasma insulin Low to zero Low, normal, or  IPF1 = MODY 4
concentration high  HNF-1β = MODY 5
Plasma C-peptide Low to zero Low, normal, or  NeuroD1/BETA2 = MODY 6
concentration high
(equal to insulin
concentration)
Genetic Yes Yes
susceptibility
Islet β cell Present Absent
abnormalities at
diagnosis
Biochemistry Hormonal Regulation of Metabolism 2 7 of 11
  Increased sensitivity of target cells to other Insulin counter
regulatory hormones
 Half-life: 8-18 minutes
A. ACTION OF GLUCAGON
 Major target organ: Liver
 Increase output by glycogenolysis and gluconeogenesis
 Ketogenesis is increased because of acetyl-CoA due to β-
oxidation of fatty acids
 Glycogenesis is decreased in glucose-dependent organs
 Proteolysis to supply glucogenic amino acids for
Figure 9. Patient with Rabson-Mendenhall syndrome [Lecturer’s PPT]
gluconeogenesis
 Increased TAGs breakdown in adipose tissue via HSL

Figure 10. Patient with Donohue syndrome [Lecturer’s PPT]

E. GESTATIONAL DIABETES MELLITUS

 Pregnancy: decrease in insulin sensitivity due to pregnancy
hormones being anti-insulin (e.g. estrogen, progesterone, hCG)
 provide adequate glucose for the fetus (to be able to extract
as much glucose as it can)
 GDM: additional decrease in insulin sensitivity in 14% of
pregnant women and inability to compensate with increase
insulin secretion
 Defect in insulin action not insulin receptor binding affinity
Figure 11. Action of Glucagon
 Patients with GDM have a 30-50% risk of developing Type II
DM after pregnancy, especially if obese Table 6. Effects on Fuel Metabolism
 Increase resistance to insulin-mediated glucose transport in LIVER ADIPOSE TISSUE
skeletal muscles TISSUE LIPOLYSIS
 Overexpression of plasma membrane glycoprotein 1 (PC- Glucose output ↑↑
1): inhibits tyrosine kinase activity by direct interaction with Ketogenesis ↑
alpha subunits  blocking insulin-induced transformation Fat synthesis
change  signaling pathway cannot proceed Gluconeogenesis ↑ – No effect
 Excessive phosphorylation of serine-threonine residues in Glycogenolysis ↑↑ ↑
the insulin receptors of skeletal muscles  down-regulation of Glycogenesis ↓
tyrosine kinase activity  no insulin signaling cascade Protein Synthesis -
 Decrease expression and phosphorylation of tyrosine
residues in IRS-1 in skeletal muscles  decrease in activity
B. SYNTHESIS OF GLUCAGON
of the signaling pathway
 2 sites of production: α cells of pancreas and L cells of
F. NEONATAL DIABETES MELLITUS intestine
 Genetic disease caused by a mutation in the KCNJ11 (K-  Precursor: Preproglucagon composed of 160 amino acids
inwardly rectifying channel subfamily J member 11) gene, which  Cleaved by a proteolase
also provides instructions for the assembly of the subunits of the  3 products: glucagon (29 AA) in α cells of pancreas, glucagon-
ATP-sensitive K+ channels of pancreatic islet cells like peptides 1 and 2 (GLP1 and 2) and glicentin-related peptide
 Leads to permanent opening of the ATP-sensitive K+ channels in L cells of intestine
→ no depolarization → no release of insulin

III. GLUCAGON
 Major counter-insulin or counter-regulatory polypeptide hormone
that is secreted in fasting and starvation state where blood
glucose is low
 Stimulation of:
 Mobilization of fuel reserves
 Fuel reserves utilized: liver glycogen during the first 24
hours, muscle glycogen for muscle use ONLY, and infinite
number of TAGs from adipocytes
 Glycogenolysis
 Gluconeogenesis
 Glucose is available to glucose-dependent tissues in between Figure 12. Synthesis of Glucagon from Preproglucagon
meals
C. STRUCTURE OF GLUCAGON
 Target organs: Liver (major) and adipocytes
 Single polypeptide chain of 29 amino acids
 No glucagon receptor in skeletal muscles

Biochemistry Hormonal Regulation of Metabolism 2 8 of 11

 Figure 13. Glucagon structure
Figure 15. Steps Involved in Glucagon Release (left) in Comparison to Insulin
Release (right)
D. GLUCAGON RECEPTOR
 G protein-coupled receptor (GPCR) in the plasma membrane I. SIGNALING CASCADE OF GLUCAGON
 With 7 hydrophobic plasma membrane spanning domains Signaling Cascade regulated by cAMP
 stimulates synthesis of intracellular second messenger, cyclic 1. Glucagon binds to receptor
adenosine monophosphate (cAMP) → activates protein kinase a. high-affinity receptors on the cell membrane (liver,
A (PKA) → phosphorylation of key regulatory enzymes muscle, adipose tissue, kidneys, other extrahepatic
E. DEGRADATION OF GLUCAGON cells; not present in muscle)
 Occurs in the liver and kidneys 2. Activate G-proteins
3. Activate adenylate cyclase; increase in cAMP synthesis from
 Cleavage of peptide bond between Ser 2 and Gln 3 from the N-
ATP
terminal
a. Mg2+ - co-factor
 Once cleaved, entire polypeptide is destroyed
b. cAMP - second messenger of glucagon
4. Activates cAMP-dependent protein kinase—Protein Kinase A
(PKA)—which has 2 regulatory (RR) and 2 catalytic (CC)
subunits
5. RR binds to cAMP and CC is released
6. CC catalyzes the phosphorylation and activation of target
Figure 14. Glucagon Linear Structure
enzymes
F. REGULATION OF GLUCAGON 7. Theses phosphorylated and activated target enzymes will
Table 7. Regulators of Release elicit the response
REGULATORS EFFECT Signaling Cascade regulated by PKA
MAJOR REGULATORS 1. Activated PKA translocates to nucleus via pores
Glucose - 2. Phosphorylates and activates cAMP-regulated proteins
Insulin - (CREB)
Amino Acids + 3. CREB then activates and binds to CBP
MINOR REGULATORS 4. Controls expression of genes with cAMP-sensitive regulatory
Cortisol + elements (CRE) on DNA
Neural input + 5. Synthesis of new proteins (These new proteins are enzymes
Epinephrine + for gluconeogenesis)
Glucagon on Carbohydrate Metabolism
G. RELASE OF GLUCAGON 1. Glucagon binds to receptor
 Released by α cells 2. Activates G-protein
 Transporter: GLUT1 3. Activates Adenylate cyclase; increase in cAMP synthesis
 Immediate source: liver glycogen 4. Activates PKA
 Liver glycogen contains glucose-6-phosphatase while muscle 5. Phosphorylation and deactivation of Phosphofructokinase 2
glycogen has NO glucose-6-phosphatase and will go through (PFK2)
glycolysis and Kreb’s cycle a. PFK2 – enzyme responsible for the activation of
 Low glucose, cannot undergo complete oxidation fructose-2,6-bisphosphatase (F26BPase)
 Mechanism: b. F26BPase – cleaves or dephosphorylates fructose-
1. Glucose is transported into the pancreatic α-cell through 2,6-bisphosphate (F26BP)
GLUT 1 6. Decreases synthesis of F26BP
2. Glucose is oxidized yielding ATP a. F26BP – normally stimulates phosphofructokinase 1
3. Increase in ATP/ADP ratio inhibits ATP gated K+ Channels, (PFK1); an allosteric effector for PFK1
causing the cell to depolarize 7. Decreased rate of glycolysis
4. Unknown transporter X will be active and cause membrane a. PFK1 – enzyme responsible for the phosphorylation
depolarization of fructose-6-phosphate (in glycolysis) to fructose-
5. Na+ channel and voltage-gated N-type and L-type Ca2+ 1,6-bisphosphate
channels are stimulated causing Na+ and Ca2+ influx 8. Increased rate of gluconeogenesis
6. Increase in calcium causes exocytosis of glucagon from I. DOWNREGULATION OF GLUCAGON
vesicles 1. Dissociation of the hormone from the receptor
2. Hydrolysis of GTP by Ga due to GTPase activity (Ga becomes
inactive)
3. Hydrolysis of cAMP via cAMP phosphodiesterase (PDE)
Biochemistry Hormonal Regulation of Metabolism 2 9 of 11
 4. Desensitization of receptors by kinases such as PKA and β-
arrestin receptor kinase (βARK)
 Phosphorylates serine residues
5. GPCRs removed from the membrane by vesicular uptake
IV. SUMMARY OF INSULIN AND GLUCAGON
 Glucose homeostasis is directed toward the maintenance of
constant blood glucose levels
 Insulin & Glucagon are the 2 major hormones that regulate the
balance between fuel mobilization & storage
 They maintain blood glucose near 80-100 mg%, despite
varying CHO intake during the day
 If dietary intake of all fuels is in excess of immediate need, it is
stored as either glycogen or TAGs. Conversely, appropriately
stored fuels are mobilized when demands require
 Insulin is released in response to CHO ingestion and promotes
glucose use as fuel and glucose storage
 Insulin secretion is regulated principally by blood glucose levels
 Glucagon promotes glucose production via glycogenolysis and
gluconeogenesis
 cAMP activates PKA, which phosphorylates key regulatory
enzymes, activating some & inhibiting others
 Hormones that antagonize insulin action, known as counter-
regulatory hormones include glucagon, epinephrine and cortisol
V. INCRETINS
 A group of metabolic hormones that stimulate a decrease in
blood glucose levels
 Released after eating & augment the secretion of insulin
released from pancreatic β cells by a blood-glucose dependent
mechanism
 Two main candidate molecules that fulfill criteria of an incretin
are the intestinal polypeptides:
 Glucagon-like peptide (GLP-1)
 Gastric inhibitory peptide (GIP; also known as glucose-
dependent insulinotropic polypeptide)
 GLP-1, GIP, Cholecystokinin & Vasoactive Intestinal Peptide
(VIP) potentiated insulin secretion
 Incretin effect: secreted by increased glucose concentration in
the gut
 GLP-1 and GIP act via GPCR:
 Increased insulin section
 (+) β cell proliferation, decreased apoptosis
 Slow the rate of absorption of nutrients by reducing gastric
emptying & may directly reduce food intake
 GLP-1 & GIP are rapidly inactivated by the enzyme dipeptidyl
peptidase-4 (DPP-4)
 Both are the members of the glucagon peptide
superfamily

VI. REFERENCES
 2021A Trans
 Lecturer’s ppt
 Bayne’s Clinical Biochemistry. 4th edition
 Lieberman, M. (2018). Mark’s Medical Biochemistry: A Clinical Approach 5th
edition
 Rodwell, V., et.al. Harper’s Illustrated Biochemistry 30th edition

Biochemistry Hormonal Regulation of Metabolism 2 10 of 11

 VII. APPENDIX

Biochemistry Hormonal Regulation of Metabolism 2 11 of 11