NUR 1205: MEDICAL BIOCHEMISTRY

METABOLISM
• The sum of the chemical reactions in an organism
Catabolism: Provides energy and building blocks for anabolism.
Anabolism: Uses energy and building blocks to build large molecules

A metabolic pathway is a sequence of enzymatically catalyzed chemical reactions in a cell

Energy Production:
Oxidation-Reduction Reactions
• Oxidation: Removal of electrons
• Reduction: Gain of electrons
• Redox reaction: An oxidation reaction paired with a reduction reaction

• In biological systems, the electrons are often associated with hydrogen atoms.
• Biological oxidations are often dehydrogenations.

The Generation of ATP

Phosphorylation:
1. Substrate level phosphorylation: transfer of a high-energy PO4– to ADP.
2. Oxidative phosphorylation: transfer of electrons from one compound to another is
used to generate ATP by chemiosmosis.

Carbohydrate Metabolism/ Utilization- Tissue Specificity

• Muscle – cardiac and skeletal
o Oxidize glucose/produce and store glycogen (fed)
o Breakdown glycogen (fasted state)
o Shift to other fuels in fasting state (fatty acids)
 • Adipose and liver
o Glucose  acetyl CoA
o Glucose to glycerol for triglyceride synthesis
o Liver releases glucose for other tissues
• Nervous system
o Always use glucose except during extreme fasts
• Reproductive tract/mammary
o Glucose required by fetus
o Lactose  major milk carbohydrate
• Red blood cells
o No mitochondria
o Oxidize glucose to lactate
o Lactate returned to liver for Gluconeogenesis

STEPS OF GLYCOLYSIS

Preparatory phase “Glucose priming”

• get glucose ready to split
o phosphorylate glucose
o molecular rearrangement
• split destabilized glucose

Pay off “Energy harvest”

• Two glyceraldehyde-3-phosphates can pass per glucose.
• Five more enzymatic reactions for a total of ten in Glycolysis. Four at this stage are
reversible.
 • Enzyme #10: Pyruvate Kinase is regulated.
• As the carbon becomes oxidized the phosphate bonds elevate in their energy potential.
• Two steps involve SLP for ATP.
Substrate-level Phosphorylation
9 O-
H2 enolase H2 C O
O O
C O P
Phosphoenolpyruvate Phosphoenolpyruvate CH2
(PEP) (PEP)

10 O-
ADP ADP
pyruvate kinase C O
ATP ATP C O
Pyruvate Pyruvate CH3

• P is transferred from PEP to ADP

o kinase enzyme
o ADP  ATP
Bioenergetics of (or Energy yield from) glycolysis:
Under anaerobic conditions:
1. Total ATP lost = 2 ATP as follows,
• One ATP in the activation of glucose to glucose-6-phosphate.
• One ATP in the activation of fructose-6-phosphate to fructose1, 6-diphosphate.
2. Total ATP gained = 4 ATP as follows,
• 2 ATP by substrate level phosphorylation from 1,3-diphosphoglycerate
• 2 ATP from substrate level phosphorylation from phosphoenol pyruvate.
3. Net ATP gained = 4 ATP gained - 2 ATP lost = 2 ATP for the anaerobic oxidation of
one mole of glucose into lactate.
Under aerobic conditions:
• Total ATP lost = 2 ATP.
• Total ATP gained = 10 ATP are generated as follows,
• 4 ATP (obtained by substrate level phosphorylation) + 2 NADH.H + chain (produced
from oxidation of glyceraldehyde-3-phosphate)  2 X 3 ATP = 6 ATP, after
oxidation in the functioning respiratory
• Net ATP gained = 8 ATP as follows,
• 10 ATP – 2 ATP = 8 ATP for the aerobic oxidation of one mole of glucose.
Glucose + 2ADP + 2Pi + 2 NAD+  2 pyruvate + 2ATP + 2NADH
Transitional phase: OXIDATION OF PYRUVATE
• Links glycolysis to the citric acid cycle
• Converts each pyruvic acid to acetyl CoA
• Step is carried out by a multienzyme complex (Pyruvate dehydrogenase) that
catalyses three reactions:
o Decarboxylation - removal of 1 C to produce acetic acid and CO2
 o Oxidation – H atoms removed from acetic acid; picked up by NAD + 
NADH + H+
o Forms acetyl CoA: Acetic acid + coenzyme A  acetyl coenzyme A (acetyl
CoA)

Central Role of Acetyl CoA

• Acetyl CoA is central to both energy production & biomolecule synthesis
• Depending on organism’s need
o build ATP
 immediate use
o build fat
 stored energy
TCA CYCLE (Krebs cycle (citric acid cycle, TCA cycle)
• Occurs twice
• Results in the oxidation of the last 4 carbon atoms
• Acetyl CoA binds with oxaloacetic acid to form citric acid
• citric acid then progresses through a series of reactions ultimately resulting in the
reformation of oxaloacetic acid

• C from acetyl CoA is oxidized creating 2 CO2 per cycle (4 total)

• One high energy phosphate compound (GTP) is produced per cycle (2 total)
• 3 NADH are produced per cycle (6 total)
 • One FADH2 is produced per cycle (2 total)
Generates many compounds available for biosynthetic purposes
• -Ketoglutarate and oxalacetate (OAA): precursors of several amino acids; OAA
also converted to phosphoenolpyruvate, a precursor of glucose
• Succinyl-CoA: required for synthesis of cytochromes, chlorophyll, and other
tetrapyrrole compounds
• Acetyl-CoA: necessary for fatty acid biosynthesis
ELECTRON TRANSPORT SYSTEMS
• Membrane associated
• Mediate transfer of electrons
• Conserve some of the energy released during transfer and use it to synthesize ATP
• Made up of 4 different proteins:
o Complexes (oxidation–reduction enzymes)
o Electron carriers with increasing reduction potential (greater affinity for
electrons).
Complexes
• Complex I (NADH dehydrogenases)
o Bound to inside surface of cytoplasmic membrane;
o Binds NADH and accepts 2 electrons and 2 protons that are passed to
flavoproteins
• Complex II (succinate dehydrogenase complex)
o Bypasses Complex I
o Feeds e and H+ from FADH directly to quinone pool
• Complex III (cytochrome bc1 complex)
o Transfers e from quinones to cytochrome c
o Cytochrome c shuttles e to cytochromes a and a3
• Complex IV: (cytochromes a and a3) - Cytochrome c oxidase
o Terminal oxidase; reduces O2 to H2O
2H+ + 2e- + ½O2 → H2O
Electron carriers
• Flavoproteins:
o Contains flavin prosthetic group (e.g., FMN, FAD) that accepts 2 electrons
and 2 protons
o Donates the electrons to the next protein in the chain
• Cytochromes
o Contain heme prosthetic groups
o Accept and donate a single electron via the iron atom in heme
• Iron - Sulfur Proteins
o Contain clusters of iron and sulfur e.g. ferredoxin
o Reduction potentials vary depending on number and position of Fe and S
atoms
o Carry electrons
 • Quinones
o Hydrophobic non-protein-containing molecules
o Accept electrons and protons but pass along electrons only

The Proton Motive Force

• ETC is oriented in the cytoplasmic membrane so that electrons are separated from
protons.
• Electron carriers arranged in order of their reduction potential
• The final carrier donates the electrons and protons to the terminal electron acceptor
• During electron transfer, several protons are released on outside of the membrane
o Protons originate from NADH and the dissociation of water
• Results in generation of pH gradient and an electrochemical potential across the
membrane (the proton motive force)
o The inside becomes electrically negative and alkaline
o The outside becomes electrically positive and acidic
Oxidative Phosphorylation
• ATP synthase (ATPase): complex that converts proton motive force into ATP

• The energy generated during oxidation (proton electrochemical gradient) is coupled to

ATP production (ADP phosphorylation)
Energetics Balance Sheet for Aerobic Respiration
Regulation (or Control) of Glycolysis
A. Key regulatory enzymes:
• Those that catalyze the irreversible steps of glycolysis and include:
1-Phosphofructokinase:
• An allosteric enzyme stimulated by high levels of fructose-6- phosphate, fructose-2,6-
diphosphate (in liver), ADP and AMP, Pi, and ammonia.
• Inhibited allosterically by ATP, low pH and citrate.
PFK-1 and F1,6-BP are reciprocally regulated

2-Hexokinase:
Accumulation of glucose-6-phosphate and inhibition of phosphofructokinase results in
accumulation of fructose-6-phosphate and glucose-6-phosphate that allosterically inhibit
hexokinase.
3-Pyruvate kinase:
• Inhibited also by excess ATP, fatty acids, and acetyl-CoA
• Stimulated by fructose-1,6-diphosphate, ADP and AMP
• It is regulated by cAMP-dependent phosphorylation-dephosphorylation mechanism

B. Hormonal regulation:
 • Insulin:
o Stimulates synthesis of glucokinase, phosphofructokinase and pyruvate kinase, so it
stimulates glycolysis.
o It also induces glucose transporters to provide cells with glucose for glycolysis.
• Adrenaline and glucagon are
• Inhibitory by inhibiting pyruvate kinase.
GLUCONEOGENESIS
• A metabolic pathway that results in the generation of glucose from non-carbohydrate
carbon substrates.
• It is one of the two main mechanisms the body uses to keep blood glucose levels from
dropping too low.
• In animals, gluconeogenesis takes place mainly in the liver.
• This process occurs during periods of fasting, starvation, or intense exercise.
Irreversible glycolytic steps bypassed
The Beginning of Gluconeogenesis

The End of Gluconeogenesis

Reciprocal regulation of glycolysis and gluconeogenesis

• The two are controlled in reciprocal fashion.
 • Two main points :
o Control PFK-1 and F1,6-BPase
o Control fate of pyruvate at PDH or Pyruvate carboxylase

• Fructose-2,6-bisphosphate (F2,6BP) is the most important allosteric regulator of

glycolysis and gluconeogenesis through its reciprocal effects on fructose1,6-
bisphosphatase and phosphofructokinase.
• Is not intermediate of both glycolysis and gluconeogenesis, a product of PFK-2, under
hormone control
• found in liver of all animals, some plants, and fungi, but not in bacteria
Gluconeogenesis from glucogenic amino acids

Gluconeogenesis from Lactate and Pyruvate (Cori's Cycle)

• Illustrating the fate of lactic acid produced by active muscles and RBCs.
• Lactate and/or pyruvate formed by the anaerobic oxidation of glucose from skeletal
muscle glycogen or glycolysis in RBCs, diffuses to blood stream and is transported to
the liver and the kidney where it is transformed into glucose by gluconeogenesis
• Pyruvate also leaves muscles as alanine after transamination particularly during long
starvation, where muscle proteins break down.
• Alanine goes to liver where transamination converts it back into pyruvate.
• The glucose formed diffuses back to the blood to be used by various tissues.
Glucose Glucose Glucose

Glucose-6-phosphate Glucose-6-phosphate

Glycogen Pyruvate Pyruvate Pyruvate Glycogen

Lactate Lactate Lactate

Alanine Alanine Alanine
Blood