Mod 5 section 2.
Enzymes: Lingual Lipase and Gastric Lipase
Location and Source:
● Lingual lipase: Secreted by glands behind the tongue.
● Gastric lipase: Secreted by mucosal cells lining the stomach.
Function:
● Both enzymes are lipases, meaning they break down lipids.
● They hydrolyze triacylglycerols (TAGs) into:
○ Diacylglycerol (DAG)
○ Monoacylglycerol (MAG)
○ Free fatty acids (FFAs)
● This process starts in the mouth and stomach, aided by mechanical grinding and
mixing.
Enzyme Properties:
● Acid-stable: Remain active in the low pH of the stomach.
● Initiate lipid digestion before pancreatic enzymes are active.
Importance in Infants:
● Critical for digesting milk fat, which is a primary energy source for infants.
● Especially important before pancreatic lipase production matures.
Clinical Relevance – Cystic Fibrosis:
● In patients with cystic fibrosis, pancreatic enzyme production is reduced.
● Lingual and gastric lipases become essential for lipid digestion in these cases.
Enzymes: Pancreatic Lipase and Cholesteryl Esterase
Location and Source:
● Secreted by the pancreas into the small intestine.
● Release is hormonally regulated in response to food entering the intestine.
Function in Lipid Digestion:
● Pancreatic lipase:
○ Breaks down triacylglycerols (TAGs) into monoacylglycerol and free fatty
acids.
○ This breakdown is crucial for absorption of fats by intestinal cells.
● Cholesteryl esterase (mentioned in text, not shown in the image):
○ Hydrolyzes cholesteryl esters into cholesterol and free fatty acids.
○ Also assists in absorbing dietary cholesterol and fat-soluble vitamins.
Purpose:
● The action of these enzymes produces smaller lipid molecules that can be absorbed
across the intestinal lining.
Visual Reaction Summary:
● TAG → (lipase + water) → DAG → MAG → monoacylglycerol + 2 free fatty acids
Key Clinical Insight:
● Without pancreatic lipase, fat digestion is severely impaired.
● This enzyme is often supplemented in patients with pancreatic insufficiency, such as
in cystic fibrosis.
Mod 5 section 3:
1. Hormone-Sensitive Lipase (HSL)
Location:
● Found in adipocytes (fat-storing cells).
Function:
● Catalyzes the release of fatty acids from stored triacylglycerols (TAGs) in fat cells.
● Converts TAGs to glycerol and free fatty acids, which can enter the bloodstream for
energy use.
Regulation:
● Activated by epinephrine (during fasting or stress, when glucose is low).
● Inactivated by insulin (when glucose is high and energy is plentiful).
2. Carnitine Palmitoyltransferase I (CPT I)
Location:
● Embedded in the outer mitochondrial membrane.
Function:
● Part of the carnitine shuttle system.
● Converts fatty acyl-CoA into fatty acyl-carnitine so it can be transported into the
mitochondrial matrix for β-oxidation (fat burning).
Regulation:
● Inhibited by malonyl-CoA, a molecule made during fatty acid synthesis.
● This inhibition prevents fatty acid degradation during times of fat synthesis, avoiding a
futile cycle.
1. Hormone-Sensitive Lipase (HSL):
● Located in adipocytes (fat cells).
● Catalyzes the hydrolysis of triacylglycerols (TAGs) into free fatty acids and glycerol.
● Works in concert with other lipases to fully release fatty acids from TAGs.
2. Another Lipase (unnamed here):
● Assists HSL by removing additional fatty acids from TAG.
● Functions through hydrolysis reactions using water.
Hormonal Regulation:
Activated by:
● Epinephrine and glucagon during low glucose levels (fasted state).
● These hormones increase cAMP through activation of adenylate cyclase.
● cAMP leads to phosphorylation and activation of HSL.
● This cascade requires ATP, but the energy used is not included in fatty acid energy
yield.
Inactivated by:
● Insulin (during high glucose levels).
● Insulin blocks the cascade by preventing adenylate cyclase activation.
1. Glycerol Kinase
● Location: Liver cells
● Function: Converts glycerol (from TAG breakdown) into glycerol phosphate, which
can then enter:
○ Gluconeogenesis to make glucose
○ Glycolysis to make energy
○ TAG synthesis (in the liver)
● Not found in adipocytes, so adipose tissue cannot re-use glycerol for fat synthesis.
Fate of Lipid Breakdown Products:
● Glycerol travels from fat cells to the liver:
○ Can be used for glycolysis or gluconeogenesis.
○ May be converted to pyruvate or glucose.
● Free fatty acids (FFAs):
○ Released from adipocytes.
○ Bind to albumin in the blood.
○ Transported to peripheral tissues for β-oxidation into acetyl-CoA, which enters
the citric acid cycle (CAC) to produce CO₂ and H₂O.
Step 2A: Activation of Fatty Acids
Enzyme: Acyl-CoA Synthetase
● Location: Cytoplasm
● Function: Converts free fatty acid + CoA + ATP → fatty acyl-CoA
● ATP is converted to AMP + PPi, which counts as 2 ATP equivalents
● This activation is required before fatty acids can be transported into the mitochondria.
Step 2B: Transport via the Carnitine Shuttle System
1. Enzyme: Carnitine Acyltransferase I (a.k.a. CPT I)
● Location: Outer mitochondrial membrane
● Function: Transfers the acyl group from fatty acyl-CoA to carnitine, forming
acyl-carnitine
● Regulation:
○ Inhibited by malonyl-CoA (to prevent simultaneous FA synthesis and
degradation)
2. Transport Protein: Translocase
● Function: Transports acyl-carnitine across the inner mitochondrial membrane into the
matrix
3. Enzyme: Carnitine Acyltransferase II (a.k.a. CPT II)
● Location: Inner mitochondrial membrane (matrix side)
● Function: Converts acyl-carnitine back into fatty acyl-CoA + free carnitine inside the
matrix, ready for β-oxidation
Key Regulatory Point:
● Malonyl-CoA (a product of fatty acid synthesis) inhibits CPT I to prevent a futile cycle
of FA degradation during FA synthesis.
Mod 3 section 4
Key Enzyme: Acetyl-CoA Carboxylase (ACC)
Function:
● Converts acetyl-CoA to malonyl-CoA
● This is the rate-limiting step in fatty acid synthesis.
Regulation:
Allosteric Control:
● Activated by citrate (signal of abundant energy and building blocks)
● Inhibited by long-chain fatty acyl-CoA (product inhibition; signals enough fatty acid is
present)
Hormonal Control:
● Activated by insulin (fed state, promotes storage)
● Inhibited by epinephrine and glucagon (fasted state)
Important Transport Step: Citrate Shuttle
● Citrate translocation from mitochondria to cytosol only occurs when mitochondrial
citrate levels are high.
● Citrate in the cytosol is converted to acetyl-CoA (via ATP-citrate lyase) and used for
fatty acid synthesis.
Energy Implication:
● This process is energy-intensive, requiring ATP and NADPH.
● Key source of NADPH: Pentose phosphate pathway and malic enzyme.
1. Citrate Synthase
● Location: Mitochondrial matrix
● Function: Combines acetyl-CoA with oxaloacetate (OAA) to form citrate
● Purpose: Allows acetyl units to be transported out of the mitochondria in the form of
citrate, since acetyl-CoA cannot cross the mitochondrial membrane
2. ATP-Citrate Lyase
● Location: Cytoplasm
● Function: Cleaves citrate (after it exits mitochondria) into acetyl-CoA and
oxaloacetate
● Requires ATP
● This reaction regenerates the cytoplasmic acetyl-CoA needed for fatty acid synthesis
Regulation Context:
● This transport only happens when mitochondrial citrate levels are high, signaling a
fed state with excess energy and substrates.
Key Enzyme: Acetyl-CoA Carboxylase (ACC)
Function:
● Catalyzes the conversion of acetyl-CoA + CO₂ + ATP → malonyl-CoA
● This is the rate-limiting and committed step of fatty acid synthesis.
● Requires ATP and biotin (as a covalently bound coenzyme)
Allosteric Regulation of ACC:
● Activated by:
○ Citrate (signals excess energy and substrates)
○ Promotes polymerization into active filaments
● Inhibited by:
○ Long-chain fatty acyl-CoA (signals sufficient fatty acid levels)
○ Promotes depolymerization into inactive dimers
Hormonal Regulation of ACC:
● Insulin:
○ Activates ACC by triggering dephosphorylation
○ Done via activation of protein phosphatases
● Epinephrine & Glucagon:
○ Inactivate ACC by triggering phosphorylation
○ Done via AMP-activated protein kinase (AMPK) or other kinases
This step is tightly controlled to ensure fatty acid synthesis only occurs during high-energy (fed)
states, and is shut down during fasting or stress.
Key Enzyme: Fatty Acid Synthase (FAS)
Function:
● A multifunctional enzyme complex responsible for the remaining steps of fatty acid
synthesis
● Builds long-chain saturated fatty acids from acetyl-CoA and malonyl-CoA
● The main product is palmitate (16:0)
Mechanism Overview:
1. Initiation:
○ One acetyl group is loaded onto the enzyme (via acetyl-CoA), becoming the
omega (ω) end
○ One malonyl group (from malonyl-CoA) is loaded for elongation, becoming the
alpha (α) end
2. Chain Elongation:
○ Each round adds a 2-carbon unit from malonyl-CoA to the growing chain
○ Repeats until the fatty acid reaches 16 carbons (palmitate)
3. Reductive Steps:
○ Each cycle consumes 2 NADPH
○ Produces CO₂ and H₂O
Key Reaction Summary:
Fatty acyl-CoA (n) + malonyl-CoA + 2 NADPH → fatty acyl-CoA (n+2) + 2 NADP⁺ + CO₂ + H₂O
Additional Notes:
● The growing acyl chain is tethered to an acyl carrier protein (ACP) within the complex.
● FAS has seven enzymatic domains, each contributing to one part of the synthesis
cycle.
1. Activation Step – Fatty Acid to Acyl-CoA
Enzyme: Acyl-CoA Synthetase
● Function: Converts a free fatty acid into fatty acyl-CoA
● Requires: CoA + ATP → AMP + PPi (uses 2 ATP equivalents)
● Purpose: Activates the fatty acid so it can be used in TAG synthesis
2. TAG Synthesis Pathway
TAGs are made from:
● 1 Glycerol-3-phosphate
● 3 Fatty acyl-CoA molecules
Enzymes involved:
● Acyltransferases: Attach fatty acids to glycerol-3-phosphate stepwise
● Phosphatases: Remove phosphate groups when needed
Glycerol-3-Phosphate Sources (Tissue-Specific):
A. Liver and Adipose Tissue:
● Glucose → Dihydroxyacetone phosphate (DHAP) → Glycerol-3-phosphate (via
glycolysis)
B. Liver Only:
● Glycerol → Glycerol-3-phosphate (via glycerol kinase)
Note:
Adipose tissue lacks glycerol kinase, so it depends on glycolysis for glycerol-3-phosphate
production and therefore needs insulin (fed state) to stimulate glucose uptake.
Key Enzyme: HMG-CoA Synthase
● Full name: 3-hydroxy-3-methylglutaryl-CoA synthase
● Location: Mitochondria of liver cells
● Function: Catalyzes the rate-limiting step in ketogenesis
● Reaction: Acetoacetyl-CoA + Acetyl-CoA → HMG-CoA
Pathway Summary:
1. Fatty acyl-CoA undergoes β-oxidation to form Acetyl-CoA
2. 2 Acetyl-CoA molecules combine to form Acetoacetyl-CoA
3. HMG-CoA Synthase converts Acetoacetyl-CoA + another Acetyl-CoA → HMG-CoA
4. HMG-CoA is cleaved to form Acetoacetate
5. Acetoacetate can be converted into:
○ Acetone (spontaneously, exhaled in breath)
○ 3-Hydroxybutyrate (reduced form, transported in blood)
During fasting, the liver produces ketone bodies (acetoacetate and 3-hydroxybutyrate) from
acetyl-CoA derived from:
● Fatty acid oxidation
● Amino acid catabolism
● Glycolysis (limited in fasting)
Key Enzyme in Peripheral Tissues: Thiophorase
Name: Succinyl-CoA:acetoacetate CoA transferase (commonly called thiophorase)
Function:
● Converts acetoacetate into acetoacetyl-CoA by transferring CoA from succinyl-CoA
● This is a critical step in using ketone bodies for energy via the TCA cycle.
⚠️ The liver lacks thiophorase, so it cannot use ketone bodies—it only
produces and exports them.
Peripheral Tissue Use of Ketones:
● 3-hydroxybutyrate is oxidized to acetoacetate (via 3-hydroxybutyrate
dehydrogenase, not shown on this slide).
● Acetoacetate is then converted to acetoacetyl-CoA by thiophorase
● Acetoacetyl-CoA → 2 Acetyl-CoA → TCA cycle → Energy (ATP)
Mod 5 section 6:
Stepwise Pathway to Cholesterol:
1. 2 Acetyl-CoA → Acetoacetyl-CoA
○ Enzyme: Thiolase
2. Acetoacetyl-CoA + Acetyl-CoA → HMG-CoA
○ Enzyme: HMG-CoA Synthase
○ Note: There are two isoforms:
■ Mitochondrial HMG-CoA Synthase → used for ketone synthesis
■ Cytosolic HMG-CoA Synthase → used for cholesterol synthesis
3. HMG-CoA → Mevalonate (cholesterol synthesis step)
○ Enzyme: HMG-CoA Reductase
○ This is the rate-limiting step in cholesterol synthesis
Key Enzyme: HMG-CoA Reductase
Function:
● Converts HMG-CoA → Mevalonate
● Uses NADPH as a reducing agent
● Rate-limiting and committed step in cholesterol synthesis
Regulation of HMG-CoA Reductase:
Hormonal Regulation:
● Activated by insulin (fed state)
● Inhibited by glucagon (fasting state)
Allosteric/Transcriptional Regulation:
● Inhibited by high cholesterol (via SREBP regulation)
● Activated by low cholesterol
Pharmacological Regulation:
● Statins are competitive inhibitors of HMG-CoA reductase
Key Enzyme: HMG-CoA Synthase
● Function: Catalyzes the formation of HMG-CoA (3-hydroxy-3-methylglutaryl-CoA)
from acetoacetyl-CoA and acetyl-CoA
● Location-specific forms:
○ Cytosolic HMG-CoA synthase → used for cholesterol synthesis
○ Mitochondrial HMG-CoA synthase → used for ketone body synthesis
Key Concept:
● The cell compartmentalizes cholesterol and ketone synthesis by using different
isoforms of the same enzyme in different cellular locations:
○ Cytosol → cholesterol synthesis
○ Mitochondria → ketogenesis
Note:
● All cells express the cytosolic form, meaning all cells can make cholesterol.
● Only liver cells express the mitochondrial form and can perform ketogenesis.
Key Enzyme: HMG-CoA Reductase
Function:
● Catalyzes the reduction of HMG-CoA to mevalonate
● Location: Cytoplasm
● Uses: 2 molecules of NADPH as reducing agents
Reaction:
3-hydroxy-3-methylglutaryl-CoA (HMG-CoA)
→ Mevalonate
● CoA + 2 NADP⁺ (byproduct)
Importance:
● This is the major control point in cholesterol biosynthesis.
● After this step, the pathway is committed to cholesterol synthesis.
Regulation of HMG-CoA Reductase:
● Activated by: Insulin
● Inhibited by:
○ Glucagon
○ Statins (competitive inhibitors)
○ High cholesterol (via SREBP pathway)
Main Enzyme Regulated: HMG-CoA Reductase
🧬 1. Transcriptional Regulation (SREBP Pathway)
● SREBP (Sterol Regulatory Element Binding Protein):
○ A transcription factor that activates the gene for HMG-CoA reductase.
○ SCAP (SREBP cleavage-activating protein) escorts SREBP from the ER →
Golgi for activation.
● Low cholesterol levels:
○ Activate SREBP, increasing transcription of HMG-CoA reductase → ↑
cholesterol synthesis.
● High cholesterol levels:
○ Inhibit SREBP processing → ↓ HMG-CoA reductase expression → ↓ cholesterol
synthesis.
⚙️ 2. Hormonal Regulation (Phosphorylation State)
● Insulin (fed state):
○ Activates protein phosphatases → dephosphorylates HMG-CoA reductase
→ activates the enzyme.
● Glucagon (fasting state):
○ Activates AMP-activated protein kinase (AMPK) → phosphorylates
HMG-CoA reductase → inactivates it.
💊 3. Drug Inhibition – Statins
● Statins are structural analogs of HMG-CoA.
● They competitively inhibit HMG-CoA reductase, reducing cholesterol synthesis.
🧬 Cholesterol Synthesis & Regulation
Step 2: Mevalonate Formation
● Reaction: HMG-CoA → Mevalonate
● Enzyme: HMG-CoA reductase
● Location: Cytoplasm
● Cofactors: 2 NADPH
● Key Point: This is the major control point in cholesterol synthesis.
🔒 Regulation of Cholesterol Biosynthesis
1. Sterol-Mediated Regulation
● Low cholesterol: Activates SREBP → ↑ HMG-CoA reductase → ↑ cholesterol
synthesis.
● High cholesterol: Inhibits SREBP → ↓ HMG-CoA reductase → ↓ cholesterol synthesis.
2. Hormonal Regulation
● Glucagon: Phosphorylates and inhibits HMG-CoA reductase → ↓ cholesterol
synthesis.
● Insulin: Dephosphorylates and activates HMG-CoA reductase → ↑ cholesterol
synthesis.
🩸 Plasma Lipoproteins Overview
Chylomicrons
● Function: Transport dietary triacylglycerol (TAG).
● Apolipoproteins: Apo B-48, C-II, and E.
● Largest, least dense lipoprotein.
Chylomicron Metabolism
● Lipoprotein lipase (LPL) hydrolyzes TAG → free fatty acids (FFA).
○ Activated by: Apo C-II.
● Insulin:
○ ↑ LPL in adipose tissue.
○ ↓ LPL in muscle (regulatory).
🧪 HDL Metabolism
● HDL function:
○ Reservoir for Apo A, C, E.
○ Transports cholesterol from peripheral tissues to the liver (reverse cholesterol
transport).
● Enzyme: PCAT (phosphatidylcholine-cholesterol acyl transferase)
○ Activated by: Apo A1.
● Key Concept: HDL = "good cholesterol" carrier, promotes homeostasis.
