KYAMBOGO UNIVERSITY

FACULTY OF SCIENCE

DEPARTMENT OF CHEMISTRY

PROGRAM: BACHELOR OF SCIENCE TECHNOLOGY –CHEMISTRY

YEAR : TWO

SEMESTER: ONE

COURSE TITLE: METABOLISM, MOLECULAR GENETICS AND BIOCHEMICAL

TECHNIQUES

COURSE CODE: SST 2102

TASK: GROUP PRESENTATION ON GLYCOGEN PHOSPHORYLASE ENZYME

REGULATION IN CARBOHYDRATE METABOLISM

No. NAME REGISTRATION SIGNATURE

NUMBER
1 NGABIRANO MEDARD 23/U/CTD/1077/PD
2 AKELLO DORINE 23/U/CTD/03493/PD
3 AMUGE RHONA OKIROR 23/U/CTD/03932/PD
4 WAKUNGA DEDONE 24/U/CTD/622/GV
5 KAINEMUKAMA MARK 23/U/CTD/06226/PD
6 ASEGE PRISCILLA 23/U/CTD/O4329/PD
7 SIGINI EMMANUEL LINO 24/U/CTD/1759/PD
8 NAMUTEBI GRACE 23/U/CTD/10414/PD
9 KIIZA SAM 24/U/CTD/469/GV
10 BALIHAMWE HILDA 23/U/CTD/05038/PD
11 NAMULONDO RACHEAL 24/U/CTD/1467/PD
 12 MUTEGEKI IVAN 23/U/CTD/08789/PD
13 AMANYA PIUS 23/U/CTD/03807/PD
14 MANZI JOEL 23/U/CTD/07858/PD
15 BUWEMBO KASHER MARK 23/U/CTD /05341/PD
16 NALWANGA ESTHER 24/U/CTD/542/GV

QUESTION

Describe and illustrate the regulation of enzyme Glycogen phosphorylase, discussing

its role in maintaining cellular energy homeostasis, and explain how changes in energy status,
hormonal signals, and allosteric effectors influence its activity.
 GLYCOGEN PHOSPHORYLASE ENZYME

Introductiong

Glycogen is the main storage form of glucose and it is stored in the liver and skeletal

muscles in the body. It serves as a reservoir of energy that can be mobilized in times of need

such as during fasting. Glycogen phosphorylase is the enzyme responsible for glycogen

breakdown in the liver and muscles. It catalyses the rate limiting step in glycogenolysis in

animals by releasing glucose-1-phosphate from the terminal alpha-1, 4-glycosidic bond. This

process is crucial for maintaining blood glucose levels during fasting and for providing

immediate energy for muscle cells during exercise.

STRUCTURE OF ENZYME GLYCOGEN PHOSPHORYLASE

Glycogen phosphorylase is a homodimer consisting of two identical subunits and has

an essential cofactor, pryridoxal phosphate (PLP), for its activity. The enzyme can be found in

two different states, glycogen phosphorylase a (GPa) and glycogen phosphorylase b (GPb). The

difference in the structures is due to phosphorylation Ser-14 residue which results in the active

form GPa. Protein phosphatases dephosphorylates the GPa to the in active form GPb. Both forms

of glycogen phosphorylase can also be found in T and R states where T is the tense form because

it appears to have a low affinity for a substrate and R is the relaxed form where it appears to have

a greater affinity for substrates.

LITERATURE REVIEW
Various advances in understanding Glycogen Phosphorylase (GP) regulation have provided

significant insights into its role in energy homeostasis, particularly under conditions of exercise

and metabolic disorders. Gonzalez et al. (2016) explored how acute and chronic exercise

influences GP activity in skeletal muscles. Their study emphasized that GP is highly responsive

to exercise-induced phosphorylation events, promoting glycogen breakdown to meet energy

demands. The findings also demonstrated that consistent physical training improves the

efficiency of these regulatory mechanisms, enabling faster GP activation during physical

exertion.

Further investigations by Meyer et al. (2018) focused on the enzyme’s allosteric regulation. This

study highlighted AMP’s role in stabilizing the R state of GP, thereby enhancing glycogenolysis

under low energy conditions. It revealed how AMP binding increases GP’s sensitivity to

phosphorylation, providing a dual regulatory mechanism where allosteric and covalent

modifications work in tandem.

Zhang et al. (2020) explored the relationship between GP dysregulation and metabolic diseases,

particularly Type 2 Diabetes. Their research identified that impaired GP activity contributes to

glycogen build-up and insulin resistance in skeletal muscle. This study brought attention to GP

as a potential therapeutic target for improving glucose metabolism and insulin sensitivity,

suggesting that controlling GP activity could alleviate some symptoms of diabetes.

Huang et al. (2022) used molecular dynamics simulations to delve deeper into GP’s structural

changes upon binding with allosteric effectors such as AMP and ATP. Their study showed how

AMP binding induces an open, active conformation of GP, facilitating glycogen breakdown,

while ATP binding stabilizes a closed, inactive conformation. This study provided a more
detailed understanding of how GP transitions between active and inactive states, illustrating the

enzyme’s structural flexibility and its relevance to metabolic control.

REGULATION OF GLYCOGEN PHOSPHORYLASE.

This regulation occurs through two primary mechanisms; allosteric regulation and covalent

modification.

Covalent modification; primarily involves phosphorylation and dephosphorylation processes of

the enzyme which switches it between active and inactive forms.

Phosphorylation; Glycogen phosphorylase b (inactive) can be converted to its active form,

phosphorylase a through phosphorylation at serine residue by an enzyme called phosphorylase

kinase. Phosphorylase itself is activated by Protein Kinase A (PKA), which is stimulated by

cyclic AMP (cAMP) signaling pathways initiated by hormones like glucagon and epinephrine.
Dephosphorylation; when blood glucose levels rise after a meal, insulin is released, activating

protein phosphatase-1 (PP1) that dephosphorylates phosphorylase a back to its less active form

phosphorylase b. This activity decreases glycogen breakdown favouring storage rather than

release of glucose.

Allosteric regulation;

This allows for a rapid response to the cell’s energy status by sensing levels of specific

metabolites.

Allosteric activation

In muscle cells when energy levels are low, AMP concentration increases. AMP acts as an

allosteric activator for phosphorylase b, promoting its conversion from the T state to the R state.

This transition enhances its activity and allows for increased glycogen breakdown to meet energy

demands.

Allosteric inhibition

When energy levels are high, glycogen phosphorylase is inhibited allosterically by ATP and

glucose-6-phosphate. These molecules bind to the specific allosteric sites on the enzyme,

inducing conformational changes that reduce its activity and decreases affinity for glycogen
substrate. ATP-mediated inhibition prevents phosphorylase kinase activation while glucose-6-

phosphate enhances ATP’s inhibitory effect.

ROLE OF GLYCOGEN PHOSPHORYLASE IN MAINTAINIG CELLULAR ENERGY

HOMEOSTASIS

Glycogen phosphorylase catalyses the release of glucose-s1-phosphate from the alpha-1,4 non

reducing ends of glycogen.

The enzyme requires a co factor, pyridoxyl phosphate (PLP) to be functional. The PLP cofactor

of GP is covalently attached to the enzyme through a Schiff base linkage with a Lysine (K)

residue.

When glycogen phosphorylase binds with glycogen, a free inorganic phosphate anion is

positioned by the PLP and the enzyme active site ,in proximity with the anomeric carbon

position of the non reducing end residue of the glycogen residue. The oxygen involved in the

glycosidic bond attacks the partially charged hydrogen associated with the phosphate ion leading

to the cleavage of the glycosidic bond. The cleaved glycogen chain leaves the active site and one

of the phosphate oxygens attacks the carbocation intermediate created during the cleavage.
This results in the release of terminal glucose residue as glucose-1-phophosphate.
HOW CHANGES IN ENERGY STATUS, HORMONAL SIGNALS AND ALLOSTERIC

EFFECTORS INFLUENCE THE ACTIVITY OF GP.

ENERGY STATUS

The cell’s energy status, is primarily indicated by the levels of ATP and AMP, directly

influences glycogen phosphorylase activity.

High Energy (High ATP Levels): When the cell has sufficient energy, ATP acts as an allosteric

inhibitor of Glycogen Phosphorylase, stabilizing the enzyme in the less active T state. This

reduces glycogen breakdown, preserving energy stores.

Low Energy (High AMP Levels): When energy is low, AMP levels rise, and AMP binds to

Glycogen Phosphorylase b, shifting it to the R state, which has a higher affinity for glycogen.

This increases glycogen breakdown, providing glucose for ATP production.

ALLOSTERIC EFFECTORS

Allosteric effectors play a crucial role in regulating the activity of glycogen phosphorylase by

influencing its conformation and consequently its ability to bind to glycogen and catalyse its

breakdown. There are two forms of allosteric effectors that is to say;

ALLOSTERIC ACTIVATORS

Adenosine Monophosphate (AMP): AMP binds to the allosteric site of glycogen phosphorylase

b. this binding induces a conformational change, shifting the enzyme from the tense state to the

relaxed state. In the relaxed state glycogen phosphorylase enzyme has a higher affinity for its
substrate glycogen. This results in enhanced glycogenolysis, allowing the rapid mobilisation of

glucose-1-phoshate for energy production.

ALLOSTERIC INHIBITORS

Glycogen phosphorylase enzyme is inhibited by Adenosine Triphosphate (ATP) and glucose-6-

phosphate.

Inhibition by ATP

When energy levels are sufficient, ATP levels rise and serves as a signal that the cell does not

require additional energy from glycogen breakdown. ATP binds to the allosteric site on glycogen

phosphorylase, stabilizing the tense state. This prevents the transition to the relaxed state and

decreases enzyme activity. As a result the affinity of GP for glycogen is reduced, leading to

decreased glycogenolysis when energy is abundant

Inhibition by Glucose-6-Phosphate (G6P)

Glucose-6-phosphate is a product of glucose metabolism, acts as an allosteric inhibitor of GP.

When glycolytic pathways are active and energy is being produced, glucose-6-phosphate levels

rise. It binds to the allosteric site of GP inducing a conformational change that further stabilises

the tense state and decreases enzyme activity.

HORMONAL SIGNALS

During fasting, the levels of glucose in blood are low, glucagon hormone is released and then it

stimulates glycogen phosphorylase activation. Glucagon binds to its receptor (G-protein

receptor), activating adenylate cyclase and increasing cAMP, which activates protein kinase A
(PKA). PKA phosphorylates and activates phosphorylase kinase which in turn phosphorylates

and activates glycogen phosphorylase hence increasing rate of enzyme activity.

During strenuous exercise, epinephrine also known as adrenaline is released, it binds to the G-

protein receptor which is related to the glucagon receptor. However, it’s specific for epinephrine

and cannot bind with glucagon. It then activates the G-protein pathway leading to protein kinase

activation which then phosphorylates and activates glycogen phosphorylase kinase which in turn

phosphorylates and activates glycogen phosphorylase thus increasing the rate of enzyme activity.

However, when sugar levels are high, insulin hormone is released into the blood, it promotes

dephosphorylation and inactivation of glycogen phosphorylase through activation of protein

phosphatase-1 (PP1) enzyme thereby decreasing the rate of enzyme activity.

Conclusion

Glycogen Phosphorylase is a central enzyme in glycogen metabolism, playing a critical role in

both glucose release and energy homeostasis. Its regulation involves complex interactions

between hormonal signals, allosteric effectors, and covalent modifications, ensuring that

glycogen is broken down only when necessary. This multi-faceted regulation allows the enzyme
to respond dynamically to changes in energy status, providing the body with the glucose it needs

for immediate and sustained energy.

REFERENCES

1. Gonzalez, J. T., et al. (2016). “Exercise-induced changes in glycogen metabolism: Role

of glycogen phosphorylase.” Journal of Applied Physiology, 120(2), 205-213.

2. Meyer, M. K., et al. (2018). “Allosteric regulation of glycogen phosphorylase: AMP

binding and its effects on enzyme kinetics.” Biochemical Journal, 475(16), 2581-2592.

3. Zhang, M., et al. (2020). “Glycogen phosphorylase and its role in type 2 diabetes:

Implications for therapy.” Diabetes Research and Clinical Practice, 166, 108290.

4. Huang, Y., et al. (2022). “Structural dynamics of glycogen phosphorylase: Insights from

molecular dynamics simulations.” Journal of Molecular Biology, 434(15), 167695.