Researching the Relationship Between

Glucagon and Energy Homeostasis

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Table of Contents
I. Introduction………………………………………………………………………………………………………………3
II. The Process of Generating Glucagon…………………………………………………………………………4
I. Summary of Proglucagon Processing……………………………………………………………….5
II. Pancreatic Post Translational Modifications…………………………………………………….6
III. The Functions of Glucagon………………………………………………………………………………………..7
I. The Role of Glucagon in Energy Homeostasis………………………………………………….8
II. Low Insulin Glucagon Ratio……………………………………………………………………………..9
III. High Insulin Glucagon Ratio…………………………………………………………………………..10
IV. Regulation of Glucagon Secretion……………………………………………………………………………11
V. Researching the Relationship Between Glucagon and Energy Homeostasis……………..12
I. Glucagon and Therapies for Type 2 Diabetes……………………………………………......13
VI. The Glucagon Amino Acid Sequence………………………………………………………………………..14
I. Glucagon and the Incretin Hormones…………………………………………………………….15
VII. Conclusion……………………………………………………………………………………………………………….16
VIII. References……………………………………………………………………………………………………………….17
IX. About ALPCO……………………………………………………………………………………………………........18

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Introduction
Glucagon is a gut hormone generated as
a result of post translational modifications
during proglucagon processing1,2,3. Research
has demonstrated that glucagon has many
physiological functions and plays a vital role
in energy homeostasis1,4.

Since glucagon directly affects insulin levels,

it has become an important biomarker in
diabetes and obesity research5. However,
specifically researching glucagon is a
challenge due to the similarities between
the hormone’s amino acid sequence and
that of other peptides4,6.

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The Process of Generating Glucagon
The process of generating glucagon
begins with the synthesis of Regulation of Preproglucagon Synthesis1,7,8,9,10
preproglucagon (PPG), the single
precursor to all of the proglucagon
peptides1. cAMP and amino acid levels

Research in rodents and humans

has shown that the synthesis of Intestinal L-cells Pancreatic α-cells Brain PPG neurons
preproglucagon is performed and
highly regulated by tissues and
cells including1,7,8,9,10:

• Intestinal L-cells1,7,8,9
• Pancreatic α-cells1,7,8,9
• PPG neurons in the solitary
tract of the brain10

The function and activity of the

promoters and inhibitors that
regulate preproglucagon synthesis Promoters Promoters Promoters
can vary based on location1,7,8,9,10. Insulin Pax6 Wnt signaling
Bowel injury MafB
Inhibitors
Inhibitors Inhibitors Wnt signaling
Wnt signaling Insulin
β-catenin Orexin-A

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Summary of Proglucagon Processing
Once preproglucagon is formed, it
undergoes a series of post translational
Post Translational Modifications During Proglucagon Processing1,7,11,12
modifications (PTMs) triggered by
different physiological, neurological,
and endocrine cues1. The first PTM Preproglucagon
generates the proglucagon molecule1.
1 180
Additional tissue, cell, and enzyme- First PTM
specific PTMs result in the production of
proglucagon peptides1,7. During this Proglucagon
process, enzymes such as prohormone
convertase 1/3 and 2 (PC1/3 and PC2) 1 160
cleave proglucagon into various active
and inactive proglucagon peptides, Tissue, cell, and enzyme-specific post translational modifications
including glucagon11,12.

Gut and brain Pancreas

PC1/3 PC2

Glucagon and other active and

inactive proglucagon peptides
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Pancreatic Post Translational Modifications
Research indicates that the enzyme PC2 is expressed at higher levels in pancreatic α-cells than compared to the gut
and brain. Therefore, the majority of glucagon is produced by α-cells1. This enzyme is responsible for processing the
proglucagon produced by the α-cells. PC2 requires the chaperone protein 7B2 in order to cleave proglucagon in the
pancreas7. The cleavages generate glucagon as well as three inactive peptide fragments: glicentin-related pancreatic
peptide (GRPP), intervening peptide 1 (IP1), and major proglucagon fragment (MPGF)1.

Proglucagon Processing in the Pancreas1,7

Proglucagon

1 160

IP-1

1 30 33 61 64 69 72 158

GRPP Glucagon Major Proglucagon Fragment

Pancreas
PC2+7B2 PC2+7B2 PC2+7B2 PC2+7B2

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The Functions of Glucagon
Organ-Specific Functions of Glucagon1,2,6,4,13,14
Research demonstrates that the
glucagon receptor (GCGR) is mainly Increases hepatic glucose
output, lipid oxidation, and
located in the liver, but is also expressed hepatocyte survival
in the kidneys, adipose tissue, pancreas, Liver
brain, and gastrointestinal tract 2,6.
Glucagon has different physiological Decreases lipid synthesis
functions depending on where the
receptor is located1,2,6,13,14.
Increases heart rate and
Heart
cardiomyocyte survival

Increases lipolysis and

thermogenesis
Adipose
tissue
Glucagon Decreases fat mass

GI tract Decreases motility

Increases glomerular
Kidney filtration and water
reabsorption

Increases satiety

Brain

Decreases food intake

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The Role of Glucagon in Energy Homeostasis
One of the most important functions
of glucagon is the role it plays in energy
homeostasis15. Glucagon works with
insulin via the insulin glucagon ratio
(IGR) which serves as a molecular
balance for regulating carbohydrate
and lipid metabolism15.

Together, glucagon and insulin keep blood vessels

blood glucose levels normalized to
ensure the right amount of fuel is
accessible15. Depending on the ratio
between glucagon and insulin, alpha β-cells
and beta cells respond differently15.

insulin

α-cells δ-cells

glucagon

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Low Insulin Glucagon Ratio
When the IGR is low, blood glucose
levels are low and glucagon is secreted
from pancreatic α-cells into the portal Low Insulin Glucagon Ratio1,2,15
vein of the liver1,2,15.

In the liver, glucagon causes blood

glucose levels to increase through
the1,2,15:
• Breakdown of glycogen into glucose
(glycogenolysis)
• Formation of new glucose
(gluconeogenesis)
• Increase in lipid oxidation and
synthesis

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High Insulin Glucagon Ratio
An influx of glucose from the liver
into the blood caused by glucagon’s
actions sends signals to pancreatic High Insulin Glucagon Ratio1,2,5,15
β-cells to release insulin, thus
increasing the IGR5.

Insulin then promotes the cellular

uptake of the newly available
glucose and suppresses glucagon
secretion. As a result, the liver stops
producing glucose and prevents the
development of a hyperglycemic
state1,2,5,15.

In addition, a high IGR supports

the biosynthesis of proteins and
reduces the formation of free
fatty acids15.

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Regulation of Glucagon Secretion
In addition to insulin and blood glucose
levels, research suggests that there are Examples of Glucagon Secretion Regulators3,16
many other regulators of glucagon
secretion including:
Type of Effect on Glucagon
Regulator Name
• Other hormones3,16 Regulator Secretion
• Metabolic factors3,16
• The nervous system3,16 Hormones Insulin

Many of the hormones that assist with Oxyntomodulin

the regulation of glucagon secretion are
proglucagon peptides, which contain GLP-1
the glucagon sequence16. Research also
demonstrates that the incretins GLP-1 GIP
and GIP regulate glucagon3.
Somatostatin

Researchers have made considerable Ghrelin

progress towards unraveling the
mechanisms behind the inhibition Leptin
and stimulation of glucagon secretion
by these regulators, however, more Epinephrine
research is still needed3,16.
Amylin

Metabolic Amino acids

Fatty acids

Glucose
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Researching the Relationship Between Glucagon
and Energy Homeostasis
Due to glucagon’s role in energy
homeostasis, the hormone is an Abnormal Glucagon Secretion and Hyperglycemia17
important biomarker in the effort to
understand diabetes. Research has
established that abnormal glucagon
secretion from α-cells plays an
important role in diabetes. Some
studies have indicated that the fasting Increased glucose
blood concentration of glucagon is formation
increased by 50% in some type 2
diabetics. The increase in glucagon Increased
concentration may lead to excessive secretion
glucose production in the liver and
contribute to hyperglycemia17.

The exact cause of increased glucagon

in type 2 diabetics has not yet been
fully explained. However, several
existing theories link the increase to:

• Irregular crosstalk between β-cells

and α-cells16 Hyperglycemia due to
• α-cell insulin resistance16 excessive glucose from liver
• Dedifferentiation of β-cells into cells
that can produce and release
glucagon16

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Glucagon and Therapies for Type 2 Diabetes
Additionally, glucagon is a target for diabetes
therapy development1. Researchers are
continuously working towards developing
new diabetes therapies that specifically target General Effects of Diabetes Drugs on Glucagon Levels5,15,16,18
glucagon and restore the functionality of α-
cells15,16. Furthermore, researchers suggest Type of Diabetes Drug Effect on Glucagon Levels
that investigating the IGR more closely may
Decreases postprandial
ultimately result in the ability to personalize DPP-4 inhibitors
secretion
diabetes treatments, as it can offer insight
when prescribing glucose lowering drugs15. Decreases postprandial
Amylin agonists
response
Dipeptidyl peptidase-4 (DPP-4) inhibitors, Reduces fasted and fed
amylin agonists, GLP-1 receptor agonists, GLP-1 receptor agonists
concentrations
and sodium-glucose cotransporter-2 (SGLT2)
inhibitors are examples of diabetes drugs that SGLT2 inhibitors Increases
have been shown to change the levels of
glucagon while restoring blood glucose levels
in individuals with T2D16. Many researchers
agree that glucagon measurement needs to be
incorporated into more study designs to gain
a better understanding of how these drugs
affect glucagon levels3,5,15.

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 The Glucagon Amino Acid Sequence
Although researchers know that including glucagon measurements in their experimental design is key to fully understanding diabetes,
glucagon’s 29 amino sequence poses challenges in the laboratory. First, the hormone’s amino acid sequence is found in several proglucagon
peptides1,3,19,20,21. Proglucagon PTMs that occur in the gut and brain produce the hormone glicentin. Glicentin can then be further cleaved
into glicentin-related polypeptide (GRPP) and oxyntomodulin1. Both glicentin and oxyntomodulin contain the glucagon sequence1. The
similar sequence homology between glucagon and these peptide hormones have created challenges in being able to specifically measure
changes in glucagon concentrations3,19,20.

Proglucagon Peptides from the Gut and Brain1,3,19,20,21

1 Glicentin 69
R S L Q D T E E K S R S F S A S A A D P L S D P D Q M N E D K R H S Q G T F T S D Y S K Y L D S R R A Q D F V Q W L M N T K R N R N N I A

1 GRPP 30 33 Oxyntomodulin 69
R S L Q D T E E K S R S F S A S Q A D P L S D P Q D M N E D H S Q G T F T S D Y S K Y L D S R R A Q D F V Q W L M N T K R N R N N I A

Proglucagon from the Pancreas

1 GRPP 30 33 Glucagon 61
R S L Q D T E E K S R S F S A S Q A D P L S D P Q D M N E D H S Q G T F T S D Y S K Y L D S R R A Q D F V Q W L M N T

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Glucagon and the Incretin Hormones
In addition to sharing sequence homology with glicentin, GRPP, and oxyntomodulin, glucagon's sequence is similar to the
incretin hormones glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP)1,6 . GLP-1 is
also derived from preproglucagon, whereas GIP is formed from prepro-glucose-dependent insulinotropic polypeptide (pre-
proGIP)22. These sequence similarities create further challenges for researchers trying to specifically measure glucagon23.

Amino Acid Sequence Similarities Between Glucagon and the Incretin Hormones GLP-1 and GIP6
1 29

Glucagon H S Q G T F T S D Y S K Y L D S R R A Q D F V Q W L M N T

GLP-1 H A E G T F T S D V S S Y L E G Q A A K E F I A W L V K G R G

1 29

Glucagon H S Q G T F T S D Y S K Y L D S R R A Q D F V Q W L M N T

GIP Y A E G T F I S D Y S I A M D K I R Q Q D F V N W L L A Q

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Conclusion
Glucagon is a gut hormone generated mainly by proglucagon processing in pancreatic α-cells1,2,3. This gut hormone has many functions
in the body including playing an essential role in energy homeostasis by acting as a counter regulatory balance to insulin via the Insulin
Glucagon Ratio (IGR)15. When blood sugar is low, glucagon is secreted and acts on the liver to promote the conversion of glycogen into
glucose for energy1,2,3,15. In addition to the IGR, metabolic factors, the nervous system, and other hormones regulate glucagon secretion3,16.

Research suggests that abnormal glucagon secretion is a key component in the development of type 2 diabetes17. Many research teams
continue working towards understanding glucagon regulation and signaling in order to develop more personalized treatments for the
disease15. However, the glucagon amino acid sequence is found within multiple other proglucagon peptides and is similar to incretin
hormones1,6. The similarities between glucagon’s amino acid sequence and that of other hormones creates challenges for diabetes
laboratories researching the relationship between glucagon and energy homeostasis1,2,3,23. Therefore, an improved method for specifically
measuring glucagon would assist researchers with uncovering the complete pathogenesis of type 2 diabetes.

Glucagon Promotion of Energy Homeostasis1,2,15

Glucagon
Low blood Blood suppressed; Energy
glucose glucose rises blood glucose homeostasis
normalizes
α-cells secrete Glycogen converted β-cells secrete
glucagon into glucose for fuel insulin

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References
1. Sandoval and D’Alessio (2015). Physiology of Proglucagon peptides: Role of glucagon and GLP-1 in health and disease. Physiol Rev 95: 513-548. PMID: 25834231.
2. Müller, et al (2017). The New Biology and Pharmacology of Glucagon. Physiol Rev. 2017 Apr;97(2):721-766. PMID: 28275047.
3. Wewer Albrechsten, et al (2016). The biology of glucagon and the consequences of hyperglucagonemia. Biomark Med. 2016 Nov;10(11):1141-1151.
PMID: 27611762.
4. Drucker and Campbell (2015). Islet α-cells and glucagon—critical regulators of energy homeostasis. Nature Reviews Endocrinology 11(6): 329-339. PMID: 25850661
5. Haedersdal, et al (2018). The Role of Glucagon in The Pathophysiology and Treatment of Type 2 Diabetes. Mayo Clin Proc. 2018 Feb;93(2):217-239.
PMID: 29307553.
6. Brubaker and Drucker (2002). Structure-Function of the Glucagon Receptor Family of G Protein-Coupled Receptors: The Glucagon, GIP, GLP-1, and GLP-1
Receptors. Receptors Channels. 2002;8(3-4):179-88. PMID: 12529935.
7. Kim and Egan (2008). The Role of Incretins in Glucose Homeostasis and Diabetes Treatment. Pharmacol Rev. 2008 Dec;60(4):470-512. PMID: 19074620.
8. Jin (2008). Mechanisms underlying proglucagon gene expression. J Endocrinol. 2008 Jul;198(1):17-28. doi: 10.1677/JOE-08-0085. PMID: 18577568.
9. Shao, et al (2013). The Wnt Signaling Pathway Effector TCF7L2 Controls Gut and Brain Proglucagon Gene Expression and Glucose Homeostasis. Diabetes. 2013
Mar; 62(3): 789–800. PMID: 22966074.
10. Trapp and Cork (2015). PPG neurons of the lower brain stem and their role in brain GLP-1 receptor activation. Am J Physiol Regul Integr Comp Physiol. 2015 Oct
15;309(8):R795-804. PMID: 26290108.
11. Holst (2007). The Physiology of Glucagon-like Peptide 1. Physiol Rev. 2007 Oct;87(4):1409-39. PMID: 17928588.
12. Blanco, et al (2014). Biochemical and cell biological properties of the human prohormone convertase 1/3 Ser357Gly mutation: a PC1/3 hypermorph.
Endocrinology. 2014 Sep;155(9):3434-47. doi: 10.1210/en.2013-2151. PMID: 24932808.
13. Habegger, et al (2010). The metabolic actions of glucagon revisited. Nat. Rev. Endocrinol. 6, 689–697. PMCID: PMC3563428
14. Tan, et al (2013). Co-administration of glucagon-like peptide-1 during glucagon infusion in man results in increased energy expenditure and amelioration of
hyperglycemia. Diabetes 62, 1131–1138. oi: 10.2337/db12-0797. PMCID: PMC3609580.
15. Kalra and Gupta (2016). The Insulin:Glucagon Ratio and the Choice of Glucose-Lowering Drugs. Diabetes Ther. 2016 Mar; 7(1): 1–9. PMID: 26965024.
16. Godoy-Matos (2014). The role of glucagon on type 2 diabetes at a glance. Diabetol Metab Syndr. 2014 Aug 24;6(1):91. doi: 10.1186/1758-5996-6-91.
PMID: 25177371.
17. D’Alessio (2011). The role of dysregulated glucagon secretion in type 2 diabetes. Diabetes Obes Metab. 2011 Oct;13 Suppl 1:126-32. PMID: 21824266.
18. Hoogwerft, et al (2008). Pramlintide, the synthetic analogue of amylin: physiology, pathophysiology, and effects on glycemic control, body weight, and selected
biomarkers of vascular risk. Vasc Health Risk Manag. 2008 Apr; 4(2): 355–362. PMID: 18561511.
19. Pacific Biomarkers (2013). Incretins and Gastrointestinal Hormones: Pathophysiology and Pre-analytical Considerations.pacbio.com.
20. Drucker Labs (2015). Oxyntomodulin. Glucagon.com
21. Pocai (2012). Unraveling oxyntomodulin, GLP1's enigmatic brother. J Endocrinol. 2012 Dec; 215(3): 335–346. PMID: 23019069.
22. Ugleholdt, et al (2006). Prohormone Convertase 1/3 is Essential for Processing of the Glucose-dependent Insulinotropic Polypeptide Precursor. J Biol Chem. 2006
Apr 21;281(16):11050-7. PMID: 16476726.
23. Wewer Albrechtsen, et al (2016). Inability of Some Commercial Assays to Measure Suppression of Glucagon Secretion. J Diabetes Res. 2016;2016:8352957. PMID:
26839899.

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against diabetes and obesity.

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Product Manager, Diabetes & Obesity
(800) 592-5726, x255
ccaillouette@alpco.com

About ALPCO
American Laboratory Products Company (ALPCO) was founded in 1991 as an importer and distributor of immunoassay
products for the North American life science markets. The company has since evolved into a premier developer and
manufacturer of immunoassay-based in vitro diagnostic reagents. Today, ALPCO offers a wide range of unique testing
solutions, providing both research scientists and healthcare professionals with vital tools for advancing research and
improving quality of care.

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