KYAMBOGO UNIVERSITY

FACULTY OF SCIENCE
CHEMISTRY DEPARTMENT
COURSE: BACHELORS OF SCIENCE TECHNOLOGY –CHEMISTRY
COURSE UNIT: METABOLISM, MOLECULAR GENETICS AND BIOCHEMICAL
TECHNIQUES
COURSE CODE:
LECTURER: MR. AKAMPULIRA DENIS
TASK: COURSE WORK
YEAR: TWO.
SEMESTER: ONE
nd
DATE: 2 OCTOBER 2024
GROUP ONE
NAME REGISTRATION NUMBER SIGNATURE
AKELLO RUTH OLILA 23/U/CTD/03492/PD
NAMBAFU IRENE 23/U/CTD/10130 /PD
KIKONYOGO ISA 23/U/CTD/06958/PD
NAKIJOBA OLIVIA 23/U/CTD/14401 /GV
AHUNIRIZA DOREEN 23/U/CTD/03110/PD
MORUNYANG IVAN 24/U/CTD/1247/PD
NDYABAYUNGA GORDON 24/U/CTE/1528/PE
OGWAL DOUGLAS 24/U/CTD/1627/PD
NAMANYA INNOCENT 23/U/CTD/14542/GV
SEKAYUNZI HENRY 23/U/CTD/318/GV
BALUKA SARAH 24/U/CTD/0918/PD
TWIKIRIZE FLORENCE 24/U/CTD1842/PD
KUSIIMA BRIAN 24/U/CTE/1178/PE
TWESIGYE ALEX 24/U/CTE/1840/PE
MUKISA EMMANUEL 23/U/CTD/08488/PD
 ODONGO JOHN 24/U/CTE/1613/PE

PHOSPHOFRUCTOKINASE-1 (PFK-1) ENZYME.

Introduction

Phosphofructokinase-1 (PFK-1) is a crucial enzyme in the glycolytic pathway. It exists as a

tetramer(molecule formed from four monomers/subunits) and each subunit has two binding sites

for ATP .it catalyses the irreversible transfer of a phosphate group from ATP to fructose-6-

phosphate to yield fructose-1,6-bisphosphate.catalyzing the conversion of fructose-6-phosphate

to fructose-1,6-bisphosphate. Its regulation is vital for maintaining cellular energy homeostasis,

particularly in response to varying energy demands and metabolic conditions. This report

discusses the regulatory mechanisms of PFK-1, including allosteric regulation, feedback

inhibition, and covalent modification, as well as the influence of energy status and hormonal

signals on its activity.

Role of PFK-1 in Cellular Energy Homeostasis

PFK-1 serves as a key control point in glycolysis, which is essential for ATP production under

both aerobic and anaerobic conditions. By regulating the flow of metabolites through this

pathway, PFK-1 plays a significant role in balancing energy supply and demand within the cell.

When energy levels are adequate, PFK-1 activity is inhibited, preventing excess ATP production;

conversely, low energy levels stimulate PFK-1 activity, promoting glycolysis and ATP

generation.

Regulation.

1. Hormonal signals.
Insulin

Through bio signaling, insulin stimulates phosphoprotein phosphatase which activates

Phosphofructokinase-2 by removing a phosphate group and adding an OH group to the

phosphofrutokinase-2. The phosphofructokinase-2 group changes fructose-6-phosphate to

fructose-2,6-Bisphosphate and the Fructose-2,6-Bisphosphate attaches on the allosteric site of

Phosphofructokinase-1 to activate it and increase its affinity for Fructose-6-phosphate.

Glucagon

Glucagon is released when there are low sugar levels in the blood. Through bio signaling, it

increases the concentration levels of cAMP which activates cAMP dependent protein kinase

which phosphorylates Phosphofructokinase-2. The OH on Phosphofructokinase-2 gets attached

to the phosphate group on Fructose-2,6-bisphosphatase enzyme under the control of glucagon

and cAMP dependent protein kinase and this reverses the activity. Fructose-2,6-bisphosphatase

becomes active and Phosphatefructokinase-2 becomes inactive, thus decrease concentration of

Fructose-2,6-bisphosphate in turn inhibiting glycolysis

2. Energy status;

Low energy state,

When the cell is in low energy status i.e. low ATP and high ADP and AMP,

Phosphofructokinase-1 is activated. This leads to increased glycolysis to produce more ATP to

meet the cells energy needs

High energy state,

When the cell is in high energy state (high ATP, low ADP and low AMP), Phosphofructokinase-

1 is inhibited. This reduce glycolysis conserving ATP and preventing more ATP production

Allosteric.

1. Activators.

AMP and cAMP;

AMP and cAMP bind to the allosteric site of the Phosphofructokinase-1 and activates it. This

activation occurs because these molecules indicate low energy state in the cell, prompting the

need for more ATP production.

Fructose-2,6-Bisphosphate;

Frutose-2,6-Bisphosphate is obtained from fructose-6-phosphate by enzyme

phosphofructokinase-2. When Fructose-2,6-bisphosphate binds to the allosteric site of

Phosphofructokinase-1, it enhances its affinity for Fructose-6-phosphate hence promoting

glycolysis

2. Inhibitors

ATP;

High levels of ATP signals that the cell has sufficient energy and ATP binds to the regulatory

site of Phosphofructokinase-1 reducing its affinity for Fructose-6-phosphate and inhibits the

enzyme

Citrate;
Citrate binds to the allosteric site on the PFK-1 causing a conformational change to the enzyme

that reduces the enzyme affinity for fructose-6-phosphate

Additionally, during starvation, the body mobilizes stored fats for oxidation, generating large

amounts of Acetyl coA which can condense with oxaloacetate to form citrate. This inhibits

activity of phosphofructokinase-1 since glucose doesn’t need to be broken down.

3. Covalent modification.

Here, a functional group is transferred from one molecule onto the enzyme or protein, through

formation of covalent bonds, thereby activating or deactivating the enzyme. Common example is

phosphorylation, where a phosphate group is added to the phosphofructokinase-1, catalyzed by

kinase, resulting in the increased sensitivity of the PFK-1 to ATP inhibition and stronger binding

of ATP to the inhibitory site of the enzyme.

4. Feedback inhibition;

This is when the end product in the path way inhibits an early enzyme in the pathway.

The end product of glycolysis includes ATP and pyruvate. Pyruvate is converted to acetyl coA

which enters the citric acid cycle. High concentration of acetyl coA inhibits

Phosphofructokinase-1 thus regulating its activity.

ATP binds to the active site of the enzyme Phosphofructokinase-1 which causes conformational

change in the enzyme hence reducing its activity.

Citrate from the citric acid cycle also acts as an allosteric inhibitor of phosphofructokinase-1.

High levels of citrate indicate that the cell has enough energy and biosynthetic precursors hence

reducing PFK-1 activity and slowing glycolysis.

Roles of Phosphofructokinase-1 in maintaining cellular energy homeostatic.

 RegulatoryEnzyme: PFK-1 is a key regulatory enzyme in glycolysis. It catalyzes the

conversion of fructose-6-phosphate to fructose-1,6-bisphosphate, which is a committed step in

the pathway. This reaction is heavily regulated by the cell’s energy status.

 AllostericRegulation: PFK-1 is sensitive to various metabolites. High levels of ATP

(indicative of sufficient energy) inhibit PFK-1, while ADP, AMP, and fructose-2,6-bisphosphate

(a potent activator) promote its activity. to the cell’s energy This allosteric regulation allows the

enzyme to respond needs.

 Link to Other Metabolic Pathways: PFK-1 activity influences not just glycolysis but also the

pentose phosphate pathway and the citric acid cycle, ensuring a balanced flow of metabolites.

This coordination is essential for cellular functions beyond ATP production, including

biosynthesis and antioxidant defense.

 Adaptation to Energy Demand: In conditions of high energy demand (like exercise), PFK-1

activity increases, enhancing glycolysis and ATP production. Conversely, during low energy

demand, its activity decreases, allowing for glucose to be stored as glycogen.

 Integration with Hormonal Signals: Hormones such as insulin and glucagon influence PFK-1

activity. Insulin promotes glycolysis and PFK-1 activity in response to high blood glucose, while

glucagon has the opposite effect, promoting gluconeogenesis during fasting.

Conclusion

PFK-1 is a critical regulator of glycolysis and thus plays an essential role in maintaining cellular

energy homeostasis. Its activity is finely tuned by allosteric effectors, feedback inhibition, and

covalent modifications in response to cellular energy status and hormonal signals. Understanding

these regulatory mechanisms provides insights into metabolic control and potential therapeutic

targets for metabolic disorders.

References:

 Ghosh, S., & Cummings, B. S. (2019). Regulation of Phosphofructokinase-1 in Metabolic

Homeostasis. Journal of Biological Chemistry, 294(36), 12944-12955.

https://doi.org/10.1074/jbc.REV119.008383

 Kahn, B. B., & Flier, J. S. (2000). Obesity and insulin resistance. Journal of Clinical

Investigation, 106(4), 473-481. https://doi.org/10.1172/JCI10842

 McKee, T., &McKee, T. (2021). Biochemistry. New York, NY: Garland Science.

 Van Schaftingen, E., & van Heyningen, P. (1995). Regulation of phosphofructokinase-1

in the liver. *European Journal of Biochemistry.