IM&N-Exam 3: Major Points

Fatty Acid Synthesis, Oxidation, and PUFAs

o Major Enzymes in Fatty Acid Synthesis + Regulation
Acetyl-CoA Carboxylase (ACC)
Catalyzes the first step of FA synthesis which is
the conversion of acetyl-CoA to malonyl-CoA uses
ATP & HCO3 Two step process requiring the use of a biotin
containing enzyme as CO2 is usedlost later
during condensation
ACC is regulated by post-translational modification
o Allosterically activated by citrate
o Phosphorylated and inhibited by AMPK/PKA
Stimulated by AMP, Glucagon,
Epinephrine
Inhibited by ATP
Enzyme is inactive under low energy
charge
o Dephosphorylated and activated by
phosphatase-2a
Regulation causes conformational changes to the
enzyme
o Active form of ACC associates in multimeric
filamentous complexes
o Inactive form of ACC is monomeric/protomer
units
Fatty Acid Synthase (FAS)
Regulated at the level of transcription
Insulin stimulates TFs such as USFs and SREBPs to
increase the expression of FAS
o PUFAs inhibit SREBPs and therefore FAS
Leptin from adipose inhibits expression of SREBP-1
as it is trying to inhibit FA synthesis and increase
energy expense
o The Carnitine Shuttle
FA entry to the mitochondria = major regulator of FA
oxidation
Carnitine shuttle involves 2 enzymes: CPT-I and CPT-II
CPT-I catalyzes transfer to FA from thiol of CoA to
hydroxyl group of carnitine (occurs in cytosol)
Antiporter channel for FA-carnitine and carnitine
CPT-II catalyzes reverse transfer of FA from
hydroxyl of carnitine to the thiol of CoA
(mitochondrial matrix)

CPT-I is inhibited by malonyl-CoA (product of FA

synthesis step 1)
Malonyl-CoA is produced by ACC which is
regulated by the mechanisms in the above section
o The Role of Malonyl-CoA in Coordination of Processes
Malonyl-CoA is the intermediate produced from ACC in
FA synthesis but also plays a role in FA oxidation
regulation
Presence of malonyl-CoA will inhibit CPT-I and therefore
block entrance of acetyl-CoA into the mitochondria
This will stall the FA oxidation process
This is intuitive, as you do not want FA oxidation and FA
synthesis occurring simultaneously (waste of energy
and metabolites)
o Fate of FA Oxidation Based on Metabolic State
Fed State
Acetyl-CoA will be oxidized via the TCA cycle so
that its electrons can feed directly into the ETC
Fasted State
Acetyl-CoA undergoes incomplete oxidation in the
liver which generates ketone bodies
o Incomplete because liver OAA is depleted as
it is being fed into GNG (cannot drive TCA
cycle)
Ketones bodies go to the tissues where they
encounter succinyl-CoA:acetoacetate-CoA
transferase which catalyzes ketone breakdown
into 2 acetyl-CoA units
o These units then feed into the TCA cycle,
which remains active in muscle, heart,
brain, etc.
Principle ketones are b-hydroxybutyrate and
acetoacetate
o Essential FAs and Deficiencies
PUFAs are essential to the diet especially linoleic acid
(omega-6) and alpha-linoleic acid (omega-3)
Other essential FAs can be synthesized from these
PUFAs
18-C PUFAs that cannot be endogenously
synthesized
Classic PUFA Deficiency
Result of a fat-free TPN leading to skin and GI
problems

Body attempts to compensate by synthesizing

omega-9s, but these PUFAs are totally useless to
the body
o High omega-9s in the urine is a good
diagnostic
Deficiency of Omega-3 PUFAs
Caused by a very high intake of omega-6s/omega3s during TPN thus allowing linoleic acid to
competitively inhibit the enzymes it shares with
alpha-linoleic acid
o Causes numbness, parathesia, and visual
blurring
Omega-3s are needed for normal brain and retinal
development as well as reducing the risk of CAD
Omega-6s are needed for membranes, PG
synthesis, LT synthesis, and use in specialized
membranes (brain, testes)
Eicosanoids and Phospholipids
o Cyclic Pathway for Eicosanoid Synthesis
Production of prostaglandins, prostacyclins, and
thromboxanes
AA is converted to PGH2 by the PG Synthase enzyme
(COX)
COX domain adds two molecular oxygen (PGG2
intermdte)
PEROX domain converts hydroperoxyhydroxyl
COX-1 is constitutive in most cells, maintains
homeostasis
COX-2 is inducible and part of the inflammatory
response
PGH2 is the precursor for synthesis of PGs, PCs, and
TBXNs
PGI synthase generates PCs (PGI2)
o Acts to vasodilate and inhibit platelet
aggregation
TXA synthase generates TBXNs (TXA2)
o Acts to vasoconstrict, promote platelet
aggregation
o Linear Pathway for Eicosanoid Synthesis
Production of leukotrienes, HETEs, and lipoxins
AA is converted to HPETEs in the first step by an isoform
of LPO
5-,12-, and 15- isoforms of LPO
o 5=LTs, 12=HETEs, 15=lipoxins

 HPETEsHETEs by glutathione peroxidase (GPX)

LTs are made from 5-HPETE which forms unstable LTA4
LTA4 is acted on by glutathione tranferase and a
glutathione-SH is added to make LTC4 (LTB4
released)
LTC4 moves out of the cell and removal of
glutamate generates LTD4 and glycine LTE4
o Transferase and dipeptidase enzymes
o Physiologic Role of Major Eicosanoids
Mediation of the inflammatory response
Production of pain and fever
Regulation of BP
Induction of blood clotting
Control of several reproductive functions (like labor)
Regulation of the sleep/wake cycles
o Functional Roles of Phospholipids
Phosphatidylserine
Used for de novo synthesis of phosphatidylcholine
PS is decarboxylated and then undergoes three
successive methylations by SAMSAH (requires
methionine)
o Methionine is deficient in many persons
diets
Phosphatidylinositol
Major molecule used in signal transduction
pathways
o PI becomes phosphorylated and is then a
good substrate for PLC which cleaves it into
IP3 and DAG
Results in increased Ca release and
PKC
Also serves as a reservoir for arachnidonic acid in
membranes which is crucial to PG synthesis
Surfactant-Dipalmitoylphosphatidylcholine
Extracellular fluid lining the lungs that decreases
surface tension in order to prevent alveolar
collapse
o Two saturate palmitate that also reduce
susceptibility for oxidation by ROS
Also comes with PGly, cholesterol, and surfactant
proteins
Deficiencies cause RDS in infants and RF is adults
(trauma)
Cardiolipin

Two molecules of phosphatidic acid held together

by glycerol esterified to the phosphate groups
o Only antigenic phospholipid
Important components of IMM and bacterial
membranes
o Serves as a proton trap and buffering agent
for the decreased pH near the IMM
Peroxidation of cardiolipins unsaturated chains
leads to formation of a CytC pore allowing it to
leak into the cytosol
o Triggers IP3Ca releasemore
CytCapoptosis
Ether Phospholipids
FA at C1 is attached in an ether linkage rather
than an ester
o Myelin contains ethanolamine plasmalogens
o Cardiomyocytes contain choline
plasmalogens
PAF is a potent mediator of the IR, anaphylactic
shock, and causes aggregation and degranulation
of platelets
o Triggers production of O2- by NTs and
MPHGs
o Activates phospholipases by triggering TM
signaling receptors leading to phospholipid
degradation
Sphingolipids
Antigenic determinants of ABO blood groups
Receptors for intracellular communication
Sphingomyelin is present in nervous tissue and
sheaths
More saturated than glycerophospholipids and
contribute to specialized very tight membranes
Ceramide is a 2nd messenger in
apoptosis/differentiation
Sphingomylein
High saturation allows it to form gel domains
called caveolae and lipid rafts that are important
for signaling, LDL-Rs, and enzymes such as eNOS
Interact with cholesterol to form liquid ordered
states
o Types of Phospholipases
PLA1: cleaves ester at FA position #1
PLA2: cleaves ester at FA position #2

PLC: cleaves phosphodiester between C3 and PO4

PLD: cleaves phosphodiester between PO4 and polar
head group
Exogenous Lipids, Lipid Trafficking/Lipoproteins, and Cholesterol
Homeostasis
o Types of Lipases
Pancreatic Lipase
Primary enzyme for the digestion of exogenous
lipids
It removes FA of all lengths from positions 1 and 3,
which generates a 2-monoacylglycerol
Assisted by another pancreatic protein, colipase
o Reduces surface tension at the lipid/water
interface and helps to anchor/stabilize the
lipase
Products from reaction are stabilized by bile salts
Lipoprotein Lipase (LPL)
Only enzyme responsible for TG hydrolysis in the
capillary
o Anchored to the endothelium via heparinSO4 PGlys
Synthesized by adipose, muscle (heart), and
lactating mammary glands before being
transported to endothelium
o Requires apoC-II for activity, inhibited by
apoC-III
Releases all three FAs from TGs into the
bloodstream and sends glycerol to the liver (for
glycolysis or GNG)
o Leftover chylomicron is now a chylomicron
remnant
Cardiac LPL is a low Km enzyme, unaltered by
metabolism
Adipose LPL is a high Km enzyme, increased in the
fed state as compared to the fasted state
Hormone-Sensitive Lipase (HSL)
Activated by phosphorylation by PKA (gluc/ep
signaling)
Primarily a DAG lipase in the breakdown of TG in
adipose
ATGL/desnutrin catalyzes the breakdown of
TAGDAG
o Induced by glucocorticoids, inhibited by
insulin

o Triglyceride Transport and Utilization

TGs are surrounded by bile salts and experience
peristalsis in the intestines so that they become
emsulfied into small droplets
Pancreatic lipase then breaks them down into 2MAG and 2 FFAs that will be absorbed via mixed
micelles
In enterocytes, FFAs will be activated to Facyl-CoAs by
fatty acyl CoA synthetases and added back to 2-MAG
(DGAT enzyme)
ApoB-48 particle then has TGs and other lipids added to
it by MTTP so that is matures into a chylomicron
Chylomicron will acquire apoCII and apoE in the blood
from HDL
ApoCII activates LPL which breaksdown the FA in
the chylomicron and releases them into the blood
FFAs are taken up by muscle for energy generation or
adipose tissue where they are reassembled into TGs for
storage
o Lipid Transport with High CHO Meals
Excess sugar has to be converted from acetyl-CoA to
FFAs using ACC and FA-Synthase (fatty acid synthesis
pathway)
DHAP is converted to gly-3-P to form the
backbone of TGs
FFAs and gly-3-P assemble into a TG using
acyltransferase enzymes and particularly DGAT
(most important)
These TGs are packaged into VLDLs with cholesterol
VLDL contains apoB-100 and apoA-I upon
excretion but will acquire apoCII and apoE from
HDL in the blood
VLDL will give some of its TGs to HDL via CETP
Most VLDL is removed by the liver, ~1/3 will be
hydrolyzed
LPL releases FFAs and glycerol converting
VLDLIDL
IDL can then be cleared at the liver of converted
to LDL
LDL is either cleared by the liver, taken up into
peripheral cells for metabolism, or converted to oxLDL
which contributes to atherosclerosis
o Cholesterol Absorption and Utilization

Exogenous cholesterol is absorbed just as exogenous

lipids are through chylomicrons
Cholesterol is not removed by LPL however and
remains with the chylomicron remnant upon
arrival at the liver
Liver takes the cholesterol and downregulates the
LDL-R and increase the circulating LDL levels
Cholesterol is transported to the tissues by VLDL which
picks up additional cholesterol from HDL and loses TGs
via LPL into the VLDL particle becomes an LDL particle
LDL binds to the LDL-R and is endocytosed into
cells
NPC1 determines the fate of cholesterol within the cell
Can be sent for incorporation into the membrane
Can be sent to the ER for esterification by ACAT
and stored within lipid droplets in the cell
LDL can sometimes encounter ROS becoming oxLDL
which is taken up by macrophages and contributes to
fatty streak formation over a long period of
timeatherosclerosis
o Regulation of LDL-R and ApoAI
LDL-R is regulated at the transcriptional level
Low cholesterol levels trigger the binding of SCAP to
SREBP
These then traffick to the Golgi where S1P and
S2P cleaves off an element that binds to the SRE
on DNA to increase txn
LDL-R can be downregulated by the actions of PCSK9
which decreases the recycling of LDL-R and targets it to
the lysosome
ApoAI is important for the process of reverse cholesterol
transport
RCT is mediated through HDL (good cholesterol)
particles
ApoAI interacts with ABCA1 and delivers
cholesterol to apoAI to from nascent (oval) HDL
particles
ABCG1 then adds additional cholesterol to nascent
HDL or lipid poor HDL3 to from mature HDL
particles
Requires the activity of NCEH cleaving CE in the
tissue
o Remodeling of HDL by LCAT and CETP
CETP

LCAT
Converts the cholesterol being pumped into HDL
by ABCA1/G1 into cholesterol esters
Cholesterol Biosynthesis and Derivatives
o Major Mechanisms of HMG-CoA Reductase Regulation
Short-term regulation includes allosteric modification,
competitive inhibition by statins, and phosphorylation of
Ser871
Low ATP, glucagon, or epinephrine trigger PKA to
phosphorylate the Ser and inactive the enzyme
Insulin triggers a phosphatase to activate the
enzyme
Long-term regulation is done at the transcriptional level
Low cholesterol causes SCAP to bind SREBP and
the two then interact with COPII for trafficking to
Golgi
o High cholesterol causes Insig to bind
SCAP/SREBP and prevents their interaction
with COPII
S1P and S2P act on SREBP in Golgi to cleave it
from SCAP and then from the membrane so that is
can diffuse and bind to SRE on DNA in the nucleus
High cholesterol will also cause Insig to bind HMGCoA reductase and reduces its long life to <1
hour
o Targets the enzyme for ubiquitination and
proteasomal degradation
o Major Metabolites in Cholesterol Synthesis Pathway
Cholesterol is used a major component of myelin, cell
membranes, is the precursor for steroids and bile acids,
is required for lipid packaging into chylomicrons, and
binds Shh for membrane insertion
7-Dehydrocholesterol is the precursor for Vitamin D
synthesis
Farnesyls and geranyls are attached to proteins in order
to imbed them within the membrane
Intermediates can be used for synthesis of dolichols,
CoQ, and isopenetyl adenine
o Regulation of Steroid Hormone Synthesis
The regulator of steroid hormone synthesis are the
trophic hormones at the substrate level and
transcription level

Transfers CEs from HDL for TGs from VLDL or LDL

particles leading to HDLs eventual catabolism

Availability of the substrate cholesterol is the primary

regulator
Conversion of cholesterol to pregeneolone by
P450scc is the first step in the synthesis of a
steroid hormones
Trophic hormones activate PKA which
phosphorylates and activates CEH increase the
availability of free cholesterol within the cell for
P450scc
PKA also phosphorylates and induces StAR protein
which moves cholesterol from the IMS to the mito.
matrix in order to make it available to P450scc
o Moves into IMS using cytoskeletal proteins
o Modulation of LDL-R by Bile Acid Pool Changes
Bile acid sequestrants are used to treat high levels of
LDL cholesterol by leading to increase cholesterol
secretion
They binds the bile acids and force them to be
excreted in the stool
This up-regulates bile acid synthesis in the liver
and decrease the intrahepatic pool of cholesterol
In response, the liver will insert additional LDL-Rs on its
membrane to replenish its supply of cholesterol
Therefore it will lower serum LDL cholesterol
levels
Adipocytes and Insulin Resistance
o Major Role of the Adipocyte in Lipid Partitioning
During the fed state, TGs are synthesized by the
adipocyte
Glycerol-3P enters the cell and has two activated
Facyl-CoA units added to it followed by removal of
its P group
DGAT then catalyzes the addition of the final
Facyl-CoA
Hormonal signaling in the fasted terms triggers lipolysis
Glucagon/epinephrine activates PKA which
phosphorylates and activates HSL
(desnutrin/ATGL)
o ATGL: TAGDAG
o HSL: DAGMAG
o Mechanism of Inflammation in Obesity
Obesity causes adipocyte to become overwhelmed by
lipids to the point that some adipocytes will die
Trigger release of adipokines/cytokines nearby

Macrophages will be attracted to the area and trigger

the inflammatory response, thus inflammation
o Defects in Lipodystrophy (LD)
LD occurs when the adipocyte cannot differentiate
leading to little or no storage of fat within the
adipocytes
Rather fat is shunted to other organs such as the liver
and muscle for storage contributing to the development
of insulin resistance and ineffective fuel swapping in the
muscle
o Insulin Signaling in the Adipocyte
A lack of insulin signaling will mobilize fuels from the
adipocyte:
Increased lipolysis and fatty acid export
Decreased lipogenesis and fatty acid esterification
Decreased lipid uptake
Mediated by inflammatory adipokines such as TNF-alpha
which will decrease protein synthesis suppressing
enzymes needed to promote storage of fat in adipocytes
o Insulin Impairment in Liver and Muscle
Liver
Inhibition of Malonyl-CoA Decarboxylase (MCD)
will increase malonyl-CoA and inhibit CPT-I
o Decrease beta-oxidation of FAs
Cytokines will induce stress kinases that prevent
the suppression of gluconeogenesis thru PEPCK
activation
Muscle
No activation of AMPK leading to no ability to
sequester TORC2 in the cytosolit moves to
nucleus and increase transcription of
gluconeogenic genes
Lack of AMPK also causes metabolic insufficiency
where the muscle cannot switch from glucose to
lipids easily