12/10/22 W4: Kreb’s cycle & glycolysis

OB1: describe the function of the TCA cycle and give it's location in the cell.

- Function: responsible for oxidizing Acetyl CoA to Carbon

dioxide, to conserve energy by transferring 8e-‘s from
intermediates to NAD+ and FAD which can enter the ETC and
make ATP. And provide produces precursors for anabolism
reduced

- Location: it occurs in the mitochondria near ETC.

/

reduced

OB2: know that NADH, FADH2, GTP, CO2 are produced in

Kreb’s cycle. reclud
reduced

3NADH + 1FADH2 + 1GTP + 2CO2

OB3: list which steps are essentially irreversible and describe the steps at which Kreb's cycle can be
controlled and the allosteric regulation involved.

- Irreversible reaction enzymes

1) citrate synthase
2) isocitrate Dehydrogenase
- Kreb’s cycle is regulated by the
3) a-ketoglutarate dehydrogenase
following mechanisms;
1. Substrate availability
- TCA cycle regulation depends on
2. Product inhibition
1) phosphorylation state of ATP
3. Competitive feedback inhibition
2) reduction state of NAD+
3) presence of Ca2+ needed for muscle contraction

- regulatory steps enzymes are the rate-limiting enzymes:

1) isocitrate Dehydrogenase
2) a-ketoglutarate dehydrogenase

- isocitrate dehydrogenase
> regulated allosterically
> has 8 subunits
> activated by ADP
> inhibited by ATP
> small change in [isocitrate] leads to large
change in rate of catalysis.

- a-ketoglutarate dehydrogenase
> not allosteric enzyme
> regulated by product inhibition by NADH
 OB4: be familiar with the function of the pyruvate dehydrogenase complex and describe how the
activity of the complex is regulated.

- PDC: links glycolysis to the TCA cycle by

oxidizing pyruvate to Acetyl CoA.

Regulation

PD KINASE PD PHOSPHOTASE PRODUCT

INHIBITION
Activation: Activation:
- Acetyl CoA - Ca2+
By binding of
- NADH Inhibition:
Acetyl CoA and
Inhibition: - when PD
NADH TO PDC
- ADP kinase is
- Pyruvate activated

OB5: compute the total energy from Kreb’s cycle in terms of ATP.

- NADH makes more ATP since it makes a larger gradient by pumping H+ in complexes I, III, IV.
FADH2 pumps H+ only in Complexes III, IV.

OB6: be familiar with anaplerotic reactions that replenish Kreb’s cycle.

- Anaplerotic reactions: are those that form

the intermediates of the metabolic
pathways.

- TCA is the center of metabolism:

Because glucose,, fatty acids, amino acids, ketone
bodies, and acetate all breakdown to form acetyl
CoA for ATP production
 Glycolysis

OB1: classify carbohydrates according to number of carbons, Aldose or ketose.

3C > triose
4C > Tetrose
5C > Pentose
6C > Hexose
7C > Heptose
8C > Octose

OB2: Define isomers, epimers, and enantiomers.

- Isomers: same empirical formula but different arrangement of atoms.

- Enantiomers: have the same chemical formula but differ in position of
hydroxyl group on one or more of their chiral carbons, can be in L or D
configurations.
- Epimers: stereoisomers that differ in only a single position of atom.

OB3: Explain the cyclization of glucose and

formation of the anomers a and B.

Cyclic structures form when carbonyl group reacts with a

hydroxyl group in the same molecule.
- oxygen becomes part of the ring
- carbonyl carbon becomes anomeric containing -OH group
- a -> OH group below
- B -> OH group above

OB4: Be familiar with the most common disaccharides and polysaccharides and know how to name their
glycosidic bonds

Disaccharides- joining of 2 monosaccharides by O-glycosidic bond. (Lactose)

Polysaccharides- joining of may monosaccharides by glycosidic bonds to form linear or branched chains.
(Starch, glycogen)
 - How to name the bond:
1. Determine if the carbons involved in the bond are anomeric carbons
or not “How many anomoric carbon in the bond? “
2. How do we know? Carbonyl group is in the ring attached to oxygen.
3. Write name of anomeric C “α or β or both” and the number of
carbons involved.

OB5: relate the function of glycolysis to its tissue and cellular location.

- glycolysis: is the metabolic pathway for the breakdown of glucose to provide energy and
intermediates for other metabolic pathways.
- Glycolysis takes place in the cytosol of all cells.
- Generates energy in form of ATP.
- does not require oxygen.

Glycolysis in tissue

Liver
Muscle RBC

- Liver maintains glucose homeostasis/ - Muscle will utilize glucose for the - glycolysis is the only way
produce intermediates to makes other production of ATP for contraction. RBC can make ATP due to
compounds “anabolic pathways “ ( glycerol, - When O2 is available, muscles use not having mitochondria.
fatty acids .... ) aerobic glycolysis to produce ATP. - anaerobic respiration.
- Prevents hyperglycemia. - When the need of energy is high
but ETC doesn’t has capacity to
produce all energy needed, the
muscle use aerobic and anaerobic
glycolysis
OB6: describe the fate of glucose under aerobic and anaerobic conditions.

- Occurs in cytoplasm
- The pyruvate is reduced to lactate
by oxidizing NADH.
- Pyruvate is not completely oxidized.
- Occurs in cytoplasm and mitochondria End product is lactic acid
- Pyruvate is completely oxidized to carbon dioxide - 2 moles of ATP are produced.
through TCA cycle.
- Cytosolic NADH is oxidized via the the shuttle.
- 32 moles of ATP are produced from one mole of glucose.
- end products carbon dioxide and water

OB7: distinguish the properties and roles of hexokinase and glucokinase

Glucokinase:
- In the liver & β-cells of the pancreas.
- glycolysis can occur in the liver even when energy
levels are high so that anabolic pathways can synthesize
energy storage compounds such as glycogen and fatty
acids
- Substrate: glucose.
- Product: glucose-6-phosphate.
- high Km & high Vmax.

Hexokinase:
- In most tissues.
- Substrate: glucose.
- Product: glucose-6-phosphate.
- low Km & low Vmax

OB8: compute the net energy yield of glycolytic pathway.

- One energy investment phase.

- one energy generation phase.
- Net gain:
1. Glucose → 2 pyruvate
2. 2 ADP → 2 ATP “ 2 used, 4 produced “
3. 2 NAD+ → 2 NADH
 OB9: list the glycosidic regulatory enzymes and their corresponding effectors

Glycolysis enzymatic regulation

Step 1 Step 2 Step 10

- most important regulatory step

- F-1,6-bisP “ A product in an earlier step
- rate- limiting step
“ acts as an allosteric activator; an
- committed step
example of feed-forward regulation.
Hexokinase - An allosteric enzyme, 4 allosteric binding sites. - Pyruvate kinase is inhibited by elevated
Glucokinase
of ATP level.

- Hyperbolic curve. - high Km; functions only

- Glucose-6-Phosphate inhibits the enzyme. when [glucose] is high.
Product inhibition - It’s high Vmax allows the Regulation by
liver to efficiently lower the energy levels “ ATP Regulation by fructose
[glucose]. & AMP” 2,6-bisphosphate
- Glucokinase (liver) is not
inhibited by G6P
- fructose 2,6-bisphosphate can be produced by
- PFK-1 is inhibited by elevated levels of ATP, which fructose 6-phosphate.
act as an “energy-rich” signal indicating an - is the most potent activator of PFK-1.
abundance of high-energy compounds. - Binding of F 2,6 BP to PFK-1 increase its
- because ATP will bind as negative homotropic affinity to F6P.
effector in allosteric site and change the shape of - able to activate the enzyme even when ATP
active site of enzyme to tense shape . levels are high “ diminished ATP effect”
- PFK-1 is activated allosterically by high
concentrations of AMP, which signal that the cell’s
energy stores are depleted.
- because AMP will bind as positive effector in
allosteric site and change the shape of active site of
enzyme to relax shape .
 - Production of ATP in glycolysis by using high-energy compounds: substrate level phosphorylation.
1. 1,3 bisphospho-glycerate & phosphoenolpyruvate are high energy compounds contain Pi group,
so they are able to give their pi group to ADP to produce ATP.
2. 4 ATP will produced, because these steps occurs twice.
3. The Pi group transferred directly from an compound to ADP.

- These three compounds

have higher negative ∆G°
than ADP or ATP, so they
are able to give their
phosphate group to ADP.
- 10% of ATP is produced
by this way

OB10: describe short and long term control of glycolysis

Long term:
- Induction/ repression.
( insulin and glucagon ).
Hormonal control

Short term:
- Allosteric.
- Covalent modification. Insulin Glucagon
( the three regulatory enzymes ).

- When plasma glucose is high, - When plasma glucose is low,

insulin stimulates: glucagon stimulates:
- gene transcription and - gene transcription and
synthesis of glucokinase, synthesis of glucokinase,
phosphofructokinase, and phosphofructokinase, and
pyruvate kinase are increased. pyruvate kinase are decreased.
- Induction - Repression.
OB11: briefly describe some clinical aspects of enzymatic deficiencies in the glycolytic pathway.

Pyruvate kinase deficiency:

- Affects red blood cells (RBC), because glycolysis is the only way to produce ATP in RBCs.
- Net gain = 0 ATP
- Reduced ATP production > Alternation in RBC membrane > **Hemolytic anemia.**

Lactic acidosis

Absence of oxygen leads to anaerobic respiration causing the buildup of lactic acid, which changes
the pH of the blood