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CARBOHYDRATE METABOLISM

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Basic Carbohydrate Structure
Glycolysis
Electron Transport Chain
Pyruvate Dehydrogenase and TCA Cycle
Glycogen Metabolism
Pentose Phosphate Shunt
Gluconeogenesis
Interconversion of Sugars
CARBOHYDRATE STRUCTURE

LACTOSE: beta-Galactose-1,4-Glucose.
1. It is a disaccharide of glucose and galactose.
2. It is considered a reducing sugar because it still has the free
aldehyde group on C1 of glucose.
3. It hooks from the 1-carbon of galactose to the 4-carbon of
glucose.

SUCROSE: alpha-Glucosyl-1,2-beta-Fructose
1. A disaccharide of glucose and fructose.

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GLYCOLYSIS / FERMENTATION

RED BLOOD CELLS: They do only glycolysis. They have no mitochondria!


NET ENERGETICS OF GLYCOLYSIS: Net +2ATP come from glycolysis. 2 ATPs
are invested in the preparatory phase, and 4 ATPs (per original glucose) are recovered
in the payoff phase.
METABOLISM AND DIFFERENT TISSUES: In nearly all tissues, glucose
is trapped in the cell by converting it to glucose-6-phosphate via the action of
hexokinase or glucokinase.
1. RED BLOOD CELLS
1. Have no mitochondria. They continually produce lactate,
which is continually excreted.
2. OH- Antiport: Lactate is continually excreted out of the
RBC, in exchange for OH- coming in. The net effect of this
is to maintain the pH by preventing it from becoming too
acidic.
2. BRAIN TISSUE
1. Needs lots of glucose. It makes no lactate -- it has lots of
mitochondria and it metabolizes all of its glucose all the
way to CO2.
3. ADIPOSE TISSUE
1. It can convert glucose-6-phosphate to glycogen

2. It has mitochondria, but not very plentiful. Most of the


glucose does not go through TCA cycle but rather is used
as building block for lipid biosynthesis.
4. MUSCLE AND HEART TISSUE
1. Can also make glycogen, but it cannot get back free
glucose -- only glucose-6-phosphate.
2. It will oxidize glucose fully as long as there is O2 around. If
the muscle runs out of oxygen, then fermentation will
take over by mass action.
3. Fermentation results in high lactate which can be
excreted and which causes muscle fatigue pain.
5. LIVER PARENCHYMAL CELL
1. Gluconeogenesis: The liver can use lactate, but first it
turns it into pyruvate.
2. Liver uses Glucokinase to trap glucose instead of
hexokinase.
3. Liver has a Glucose-6-Phosphatase that can actually
restore free glucose, which it then can secrete into the
blood. The liver is the major source of blood-glucose
secretion.
4. If the liver is making fat, then that is a sign of poor health.
GLUCOKINASE -VS- HEXOKINASE:
1. Glucokinase has a higher km! It has a lower than normal affinity
for trapping glucose in the liver.
1. This means it has a higher saturation point and a higher
capacity for trapping glucose via the kinase activity. This
allows the liver to act as a buffer in taking up extra blood
glucose.

2. Hexokinase is inhibited by its product, G-6-P, whereas


glucokinase is not. This again allows the liver to take up
maximal amounts of glucose.
3. Glucokinase is inhibited by F-6-P and F-1-P, whereas
hexokinase is not.
CORI CYCLE:
1. Muscle: Glucose ------> Lactate via glycolysis.
2. Lactate goes to the liver, where it is taken up.
3. Liver: Lactate ------> Glucose via gluconeogenesis
INVESTMENT PHASE OF GLYCOLYSIS: Steps 1 thru 3. It requires 2 molecules of
ATP.
Overall: 1 Glucose ------> 2 Glyceraldehyde-3-Phosphate.
1. Glucose + ATP ------> Glucose-6-Phosphate + ADP.
Phosphorylation of Glucose.
1. Hexokinase catalyzes the addition of phosphate from
ATP to glucose. It works on other hexose sugars too. It is
an induced fit.
2. Intermediate:
1. Mg+2 is required as a cofactor in the substrate. The
true phosphorylating agent is
the MgATP2- complex.
2. Intermediate is an induced fit.
3. First ATP comes from here.
2. Glucose-6-Phosphate ------> Fructose-6-Phosphate.
1. Reversible, catalyzed by phosphohexose isomerase.
2. Changing a hexose to a ketose.

3. Fructose-6-Phosphate + ATP ------> Fructose-1,6biphosphate (FBP) + ADP


1. Catalyzed by phosphofructokinase (PFK). This the
primary regulatory enzyme.
1. Inhibited when there is an excess of ATP.
2. Second ATP comes from here.
1. THIS IS IRREVERSIBLE -- THE COMMITTED STEP
FISSION STAGE: Stages 4 and 5 (or just 4, depending on how you'd like to divide it
up).
1. Fructose-1,6-biphosphate ------> Glyceraldehyde-3Phosphate + Dihydroxyacetone Phosphate -- Cleavage of
Fructose-6-Phosphate.
1. Catalyzed by Fructose Biphosphate Aldolase. It is a
reversible aldol condensation.
2. Dihydroxyacetone Phosphate <====> Glyceraldehyde3-Phosphate
1. So now we have made two glyceraldehyde phosphates.
These guys are in reversible equilibrium with each other,
but the reaction is normally driven toward Glyc-3Phosphate by mass action, i.e. that product is quickly
taken up in the next step.
2. The reversible equilibrium is catalyzed by triose
phosphate isomerase.
THE PAYOFF PHASE: Steps 6 thru 10. Now that we have two molecules of
Glyceraldehyde-3-Phosphate, all of the following occur twice per molecule of
glucose.
1. Glyceraldehyde-3-Phosphate + Pi + NAD+ ------> NADH +
1,3-bisphosphoglycerate -- Oxidation and phosphorylation.

1. Catalyzed by Glyceraldehyde-3-Phosphate
Dehydrogenase.
2. An aldehyde is oxidized to a carboxylic acid.
3. An inorganic phosphate group is attached.
4. NAD+ ------> NADH -- It is a redox reaction, coupled to
reduction of NAD+ to NADH.
5. The product is a an "acyl phosphate" -- a type of
anhydride. It has very high phosphorylation potential.
2. 1,3-bisphosphoglycerate + ADP ------> 3Phosphoglycerate + ATP
1. Reaction is "coupled" with #6. Bisphosphoglycerate is the
common intermediate.
2. Produces *two* ATP per original glucose. This is the
BREAK-EVEN POINT. We have recovered our original
investment.
3. Very exergonic (DeltaG is even more negative than that
required for ADP ------> ATP). The product is quite stable.
4. Remember that 1,3-bisphosphoglycerate is a powerful
phosphorylating agent.
5. Catalyzed by phosphoglycerate kinase.
6. This is referred to as substrate-level phosphorylation,
via a coupled reaction with #6, as opposed to oxidative
phosphorylation, via electron transport gradients.
1. Or, it is just a direct transfer of phosphate from one
compound to another.
3. 3-Phosphoglycerate ------> 2-Phosphoglycerate. Simple
isomerization.
1. Catalyzed by phosphoglycerate mutase.

2. Intermediate: 2,3-Bisphosphoglycerate (2,3-BPG).


1. 2,3-BPG decreases hemoglobin affinity for oxygen.
2. This step in the reaction can serve as a source for
2,3-BPG in the cell, in red blood cells, e.g.
3. When the phosphoglycerate mutase enzyme is
bound to the membrane, it is partaking in glycolysis,
but when it is set free in the cytosol, it can give you
free 2,3-BPG to be used for other purposes.
4. The enzyme can also function as a 2,3-BPG
Phosphatase, working in reverse.
4. 2-Phosphoglycerate ------> Phosphoenolpyruvate (PEP).
Dehydration reaction.
1. PEP has a high phosphorylation potential (much higher
than reactant).
2. Enzyme is enolase. Simple beta-removal of water.
5. Phosphoenolpyruvate + ADP ------> Pyruvate + ATP
1. The substrate is now completely dephosphorylated.
2. 2 more ATPs produced per original molecule of glucose.
3. Catalyzed by Pyruvate Kinase. Another substrate-level
coupling reaction.
ENERGETICS OF GLYCOLYSIS: Total energy yielded per original molecule of
glucose.
1. +2 NADH: Glyceraldehyde-3-Phosphate Dehydrogenase (step
6)
2. +2 ATP net
1. -1 ATP (per original glucose): Hexokinase (step 1)

2. -1 ATP (per original glucose): Phosphofructokinase (step


3)
3. +2 ATP (per original glucose): Phosphoglycerate
Kinase (step 7)
4. +2 ATP (per original glucose): Pyruvate Kinase (step 10)
LACTIC ACID FERMENTATION: In the absence of oxygen, it allows for the
NADH to be reoxidized to NAD+. This is considered the official "end" to glycolysis,
in the absence of oxygen.
1. REDOX Reaction: One step process.
1. REDUCTION: Pyruvate is reduced to lactate. (Keto
changed to 2 alcohol).
2. OXIDATION: NADH is oxidized to NAD+
2. Note that overall process from Glucose to Lactate involves
no net change in the oxidation state of carbon. It is the same in
glucose as in lactate.
3. No ATP produced!!! The purpose is to regenerate the NAD+.
4. Catalyst is lactate dehydrogenase (LDH). Note that
although it is named dehydrogenase, this reaction is actually a
hydrogenation (hydrogens are added). Hence the enzyme is
named for the reverse reaction.
1. This enzyme is used in REVERSE, too. Both reactions are
essential:
1. Pyruvate ------> Lactate: Gives us NAD+ for glycolysis
and TCA cycle.
2. Lactate ------> Pyruvate: Gives us NADH for electron
transport chain, and restores pyruvate for further
metabolism.
3. THE FATE OF LACTATE -- Ultimately the ONLY thing it
can do is go back to pyruvate! It can then be

resynthesized back to glucose via gluconeogenesis,


or pyruvate can be oxidized via pyruvate
dehydrogenase.
REGULATION OF GLYCOLYSIS:
1. Metabolic Crossover: The notion that all intermediates
before an inhibited step will tend to build up, while all those
intermediates after the step will tend to decrease.
2. PASTEUR EFFECT: Glycolysis is substantially inhibited under
aerobic conditions.
1. The step that it is inhibited at is
the Phosphofructokinase (PFK) step of Fructose-1Phosphate ------> Fructose-1,6-Biphosphate
2. O2 makes LDH work in reverse -- and fast -- to make
pyruvate to be used in the TCA cycle.
3. NON-PHYSIOLOGICAL controllers:
1. 2-Deoxyglucose -- No OH-group at 2'-position results in
significant inhibition of glucose.
2. Sulfhydryl Reagents (Iodoacetic acid) -- they ruin
glycerol phosphate dehydrogenase, which has an
essential sulfhydryl group.
3. Fluoride Ion -- Potent inhibitor of enolase.
1. The concentration required to inhibit glycolysis is
orders of magnitude higher than that found added in
water or toothpaste.
4. Arsenate -- phosphate analog will form 1-arsenate-3phosphoglycerate which then spontaneously
hydrolyzes, to ruin glycolysis at the glyceraldehyde-3phosphate dehydrogenase step.
4. REGULATORY ENZYMES:

1. Hexokinase is inhibited by its product, Glucose-6Phosphate.


2. Phosphofructokinase (PFK) generally is inhibited by
energy-rich indicators and is stimulated by things that
indicate a need for energy.
1. Stimulated by: AMP, inorganic phosphate, Fructose2,6-biphosphate
2. Inhibited by: ATP, citrate, low pH (i.e. high levels of
intracellular lactate).
5. Fructose-2,6-biphosphate: A key regulating enzyme. Its
levels are responsive to the effects of insulin and glucagon.
1. ACTION: It increase the rate of glycolysis.
2. REGULATION: It is stimulated by insulin and inhibited by
glucagon, which makes sense if you think that insulin
promotes the uptake and utilization of sugars.
FUTILE CYCLES -- generation of heat
1. You can go back and forth between Glucose <====> Glucose6-Phosphate, with the net effect being the loss of ATP as heat.
A different enzyme catalyzes it in each direction. This is called
a futile cycle.
2. Malignant Hyperthermia -- an adverse reaction to Halothane
Anesthesia in a small percentage of people, leading to high
fever and potentially death.
OXIDATIVE DECARBOXYLATION / ELECTRON-TRANSPORT CHAIN

HIGH ENERGY COMPOUND STRUCTURE


1. ATP
1. Its anhydride bonds have a free energy of hydrolysis
of 7.3 kcal / mol,. which is therefore designated as the
cutoff point for a high-energy compound.

2. Almost always found as a magnesium salt. Mg+2 helps


to neutralize the negative charges of the phosphate.
2. 3'-5' Cyclic AMP (cAMP)
1. Has no high energy anhydride bond, but rather has a
high-energy strained ring structure.
2. Phosphodiesterase cleaves cAMP ------> 5'-AMP. The
equilibrium actually favors 5'-AMP.
3. Caffeine inhibits phosphodiesterase, keeping cAMP levels
high.
3. ADP -- no natural reactions use ADP as an energy source,
despite that it has a reactive anhydride bond.
1. ADP is utilized by converting it back to ATP, via the
reaction: 2ADP ------> ATP + AMP
4. 1,3-Bisphosphoglycerate
5. Acetyl Phosphate
6. Creatine Phosphate
1. Found in skeletal muscle. It has a phosphoramide bond (PO-N)
2. Rapidly working muscles: Creatine Phosphate supplies
depleted ADPs with a phosphate group, to recharge them
back to ATP.
3. Creatine Phosphate is excreted in the urine at a constant
rate proportional to muscle mass. Measuring CP in the
urine is a reliable indicator of muscle mass.
7. Phosphoenol Pyruvate
8. COENZYME-A: It has the following groups
1. An ADP base.

2. A Pantothenic Acid group, which must be supplied by


the diet.
3. A mercaptoethylamine group.
GLYCEROL-PHOSPHATE SHUTTLE: TRANSFERS NADH FROM CYTOSOL
TO MITOCHONDRIA.
1. It results in production of FADH2 on the other side, which enters
electron-transport chain in the middle, thereby yielding only 2
ATP.
2. Mechanism:
1. OUTSIDE Membrane:
1. Oxid: NADH ------> NAD+
2. Red: Dihydroxy-Acetone Phosphate (DHAP) ------>
Glycerol-3-Phosphate.
1. Note this is NOT the same as the
Glyceraldehyde-3-Phosphate in glycolysis.
2. The glycerol-3-phosphate then traverses the membrane.
3. INSIDE Membrane:
1. Oxid: Glycerol-3-Phosphate ------> DHAP
2. Red: FAD ------> FADH2
MALATE SHUTTLE: TRANSFERS NADH FROM CYTOSOL TO
MITOCHONDRIA
1. It results in production of NADH on the other side, which enters
at the beginning of the electron-transport chain, thereby
yielding 3 ATP.
2. Mechanism:
1. OUTSIDE Membrane

1. Oxid: NADH ------> NAD+


2. Red: Oxaloacetate ------> Malic Acid
2. Malate then traverses the mitochondrial membrane.
3. INSIDE Membrane
1. Oxid: Malic Acid ------> Oxaloacetate
2. Red: NAD+ ------> NADH
ELECTRON CARRIERS -- STRUCTURE
1. NADH: Nicotinamide Adenine Dinucleotide
1. Has a Nicotinamide Ring that carries the active carbon
that is alternatively oxidized and reduced.
1. The ring is AROMATIC when oxidized and is NONAROMATIC (in a cyclohexodiene form) when reduced.
2. We derive the nicotinic acid from our diet, from niacin.
2. FADH2: Flavin Adenine Dinucleotide
1. Derived from vitamin riboflavin
2. Has a unique 3-ring isoalloxazine structure, in which the
two active hydrogens are found.
3. Contains a ribitol alcohol group.
4. FMN is similar to it.
THE ELECTRON TRANSPORT CHAIN: It occurs on the outer surface of the inner
mitochondrial membrane, through a series of membrane-boundelectron-shuttles that
pass electrons from one complex to the next without letting go of intermediates.
1. Enzymes involved:
1. Flavoproteins

2. Often contain Non-Heme iron, which interchanges


between Fe+2 and Fe+3 forms.
2. STEPS: Successive oxidations and reduction are approx. as
follows:
OXIDATION

REDUCTION

DESCRIPTION

NADH ------> NAD+

FMNH2 ------> FMN

This first step is a two


electron transfer

FMN ------> FMNH2

Fe+3 ------> Fe+2

From here forward, only


one electron at a time is
transferred

Fe+2 ------> Fe+3

Quinone ------>
Hydroquinone

Coenzyme-Q contains
the quinone group. THIS IS
THE MIDDLE STEP, WHERE
FADH2 IS ADDED IF
PRESENT

Hydroquinone ------>
Quinone

Fe+3 ------> Fe+2

Cytochrome contains the


Fe this time, and various
cytochromes contain
heme- irons for the next
several steps.

CYTOCHROMES: They are heme-containing proteins similar to hemoglobin.


Different cytochromes carry the electrons for the next several steps.
1. Each time exchanging Fe(II) and Fe(III), we have: Cytochrome
b-566 ------> Cytochrome b-562 ------> Non-heme FeS protein
------> Cytochrome C1 ------> Cytochrome c ------> Cytochrome
a a3.
2. Cytochrome-a a3 is what finally reacts with oxygen to yield
H2O
ATP SYNTHESIS AND PROTON PUMPING:
1. Protons are pumped from the inside of the mitochondrial
matrix outside, to the intermembrane space.

2. They then flow back into the matrix by an electrochemical


gradient, and in so doing they catalyze the synthesis of ATP by
an ATPase working in reverse.
3. The ATPase is integral to the inner mitochondrial membrane.
4. pH Gradient: The electrochemical gradient can also be looked
at as a pH gradient, where the matrix is more basic and
the intermembrane space is more acidic.
SUPEROXIDE ANION: O2-, a harmful partially reduced form of O 2 (i.e. a peroxide
derivative). This is very reactive and harmful.
1. We break it down as a defense mechanism, by a two-step
process. This occurs in the organelles, peroxisomes.
1. Superoxide Dismutase: O2- ------> H2O2, aka hydrogen
peroxide
2. Catalase: H2O2 ------> H2O + O2
2. CLINICAL -- newborns are sensitive to high oxygen tension,
because they have not yet developed superoxide dismutase.
1. The retinal tissue is what is affected. Blindness may
result.
3. CLINICAL -- these oxidative processes are thought to play an
important role in aging.
UNCOUPLERS: Agents that decrease the efficiency of the electron-transport ATPsynthase gradient, causing a lower production of ATP and a greater amount of heat
given off.
1. Keep in mind that the production of ATP in the electron
transport chain is not stoichiometric. There is no 1:1
relationship, and the yields are approximate amounts based on
energetic calculations.
2. Dinitrophenol (DNP): A well-known uncoupling agent. It
would make a good diet pill as an uncoupling agent (consume

energy and give it off as heat rather than make ATP), except it
has terrible side effects -- severe cataracts.
3. Thyroxine = the body's natural uncoupling agent. Thyroxine is
released in response to cold temperature to raise body
temperature by acting as an uncoupler in the electrontransport chain.
PYRUVATE DEHYDROGENASE + THE TRICARBOXYLIC ACID CYCLE

OVERALL REACTION: The reaction occurs in the mitochondria.


1. Pyruvate ------> Acetyl-CoA + CO2
2. Multi-enzyme complex, the intermediates remain bound to the
enzyme. Enzyme channeling.
3. Overall reaction is irreversible, a REDOX Reaction.
1. OXID: Pyruvate ------> Acetyl-CoA
2. RED: NAD+ ------> NADH
COFACTORS REQUIRED FOR PYRUVATE DEHYDROGENASE: Five different
coenzymes are in the reaction:
1. Thiamine pyrophosphate (TPP)
1. From vitamin Thiamin
2. Same as that which catalyzes Decarboxylation of
Pyruvate ------> Lactic Acid in Fermentation.
3. It has an unusual 5-membered heterocyclic ring,
containing a sulfur.
2. Flavine Adenine Dinucleotide (FAD)
1. From vitamin Riboflavin
3. Coenzyme-A (CoA)
1. From vitamin pantothenate

4. Nicotinamide Adenine Dinucleotide (NAD+)


1. From vitamin niacin
5. Lipoate: Has two thiol (SH) groups.
1. Oxidation of Lipoate yields two disulfide (S-S) bonds.
2. Reduced Form: (SH) Lipoate acts as an acyl carrier.
3. Oxidized form: (SS) Lipoate acts as an electron carrier.
4. Lipoate is usually found bound to a Lysine of the enzyme
it is a cofactor for.
Coenzyme-A: Has a reactive Thiol (SH) group.
1. When acyl groups get hooked on (i.e. acetyl), they form a
thioester with very high free energies of hydrolysis.
PYRUVATE DEHYDROGENASE COMPLEX (PDC): The enzyme complex that
catalyzes conversion of pyruvate to Acetyl-CoA.
1. (E1) PYRUVATE DEHYDROGENASE:
1. COFACTOR: TPP (Thiamine Pyrophosphate)-- Thiazolium
ring. It acts as an electron sink -- it can take electrons into
its ring and stabilize them.
2. (E2) DIHYDROLIPOYL TRANSACETYLASE:
1. COFACTORS: Lipoate, CoA
2. The Core: E2 sits between E1 and E3.
1. Lipoate: Its long chain hooks to a Lys residue on the
enzyme.
2. It has oxidized (disulfide) and reduced (-SH, -SH)
forms, and can act both as an acyl carrier and an
electron carrier.

3. Lipolysyl Groups: The name of the long chains at


the end of the lipoate, which are hooked to Lys
residues on the enzyme.
3. (E3) DIHYDROLIPOYL DEHYDROGENASE:
1. COFACTORS: FAD, NAD
1. FAD: The reactive part are the two CN bonds in the
center of the ring.
OXIDATIVE DECARBOXYLATION (Pyruvate Dehydrogenase): FIVE STEPS,
occurs through substrate channeling. The enzyme complex never leaves the substrate!
1. Step 1: Pyruvate (3C) decarboxylates to 2 Carbons
1. Enzyme: Pyruvate Dehydrogenase (E1)
2. CO2 comes off -- this is the FIRST CO2 that comes off from
the original glucose.
3. The remaining C=O group turn into a CH2OH group, and it
hooks to TPP.
2. Step 2: 2-Carbon-Alcohol oxidizes to carboxylic acid
(Acetate)
1. OXID: Alcohol ------> Carboxylic Acid
2. RED: Lipoyl disulfide (-S-S-) group is reduced to 2 Thiols
(2 -SH).
3. Step 3: Esterification of the Acid with the Lipoyl group.
1. Step 3a: Acetate is esterified to one of the Lipoyl -SH
groups (to form SCH3CO2).
2. Step 3b: The Ester from (a) then transesterifies with
CoA-SH (The CoA has an SH on it!) to form Acetyl-CoA,
which is given off.

3. Enzyme: Dihydrolipoyl Transacetylase -- This is the


enzyme that yields Acetyl-CoA.
4. Step 4: Lipoyl group is reoxidized
1. OXID: Lipoyl-SH ------> Lipoyl-Disulfide (-S-S-)
2. RED: FAD ------> FADH2
3. Enzyme: Dihydrolipoyl Dehydrogenase
5. Step 5: FAD is regenerated (reoxidized)
1. OXID: FADH2 ------> FAD
2. RED: NAD ------> NADH
REGULATION OF PYRUVATE DEHYDROGENASE:
1. Pyruvate Dehydrogenase Kinase (PDH-Kinase) uses ATP
to phosphorylate Pyruvate Dehydrogenase to inactive it. So,
the activity of PDH is controlled by phosphorylation.
1. PHOSPHORYLATED FORM: Phosphopyruvate. INACTIVE
2. Pyruvate Dehydrogenase Phosphatase (PDHPhosphatase) dephosphorylates it.
1. DEPHOSPHORYLATED FORM: Pyruvate Dehydrogenase.
ACTIVE.
2. The phosphatase is active all the time, so the ultimate
phosphorylation control then depends on the levels of the
PDH-Kinase present.
3. POSITIVE CONTROLS on the kinase: These things turn off
production of PDH. They are the products of PDH itself
-- Acetyl-CoA and NADH.
1. Increase Kinase Activity ------> Decrease PDH activity -----> Less Acetyl-CoA is made.

4. NEGATIVE CONTROLS on the kinase: These things turn on


production of PDH. They are the substrates of the PDH reaction
-- Pyruvate,NAD+, and Coenzyme-A.
1. Inhibit the kinase ------> Maintain PDH activity ------> More
Acetyl-CoA is made.
OVERALL ENERGETICS OF THE TCA CYCLE: You get an energetic equivalent
of 12 ATP per turn (i.e. 24ATP per original glucose) of the cycle, and 36 ATP total
when you include Glycolysis.
Item

ATP Value

Enzymatic Reaction

2 NADH

6 ATP

Glyceraldehyde-3-Phosphate
Dehydrogenase (glycolysis)

2 ATP

2 ATP

Phosphoglycerate Kinase (glycolysis)

1 NADH

3 ATP

Isocitrate Dehydrogenase (TCA: form alphaKetoglutarate)

1 NADH

3 ATP

alpha-Ketoglutarate Dehydrogenase (TCA: form


Succinyl-CoA)

1 NADH

3 ATP

L-Malate Dehydrogenase (TCA: form


Oxaloacetate)

1 FADH2

2 ATP

Succinate Dehydrogenase (TCA: form


Fumarate)

1 GTP

1 ATP

Succinate Thiokinase (TCA: form Succinate)

TOTAL:

8 ATP per glucose from


glycolysis.

12 ATP per turn of TCA


Cycle, or 24 ATPper
glucose from TCA cycle.
GRAND
TOTAL:

38 ATP per original


glucose

ACETYL-COA:
1. Acetyl-CoA cannot lead to sugar.
2. Sources of Acetyl-CoA
1. Glucose (via pyruvate)
2. Fats (via beta-Oxidation)
3. Proteins
3. Fates of Acetyl-CoA
1. TCA Cycle
2. Fatty Acid / Sterol Synthesis
3. Ketone Bodies
STEPS OF THE TRICARBOXYLIC ACID CYCLE
1. STEP I: Acetyl-CoA + Oxaloacetate ------>
Citrate + CoEnzyme-A -- THE COMMITTED STEP
1. (2C + 4C ------> 6C)
2. Condensation reaction.
3. ENZYME: Citrate Synthase.
4. INTERMEDIATE: Citroyl-CoA, which does not detach from
enzyme. Then, CoA comes off to yield Citrate + CoA.
5. CoA has the thioester linkage with high free-energy of
hydrolysis. This makes the forward reaction favored and
drives essentially the whole process.
6. CoA is then recycled back to the Oxidative
Decarboxylation system.
7. Equilibrium in this reaction very much favors citrate.

2. STEP II: Citrate ------> Isocitrate


1. ENZYME: Aconitase.
1. It contains an iron-sulfur center (prosthetic group)
2. INTERMEDIATE: cis-Aconitate (2 3C intermediates). Does
not dissociate from enzyme.
3. H2O can be added to cis-aconitate in two ways. One leads
to citrate and the other leads to isocitrate.
4. Equilibrium reaction is driven to right by quick removal of
isocitrate in next reaction.
3. STEP III: Isocitrate ------> alpha-Ketoglutarate + CO2
1. Oxidative Decarboxylation. Now we have a 5C compound.
2. ENZYME: Isocitrate Dehydrogenase.
3. Concurrent Reduction: NAD+ ------> NADH
4. Intermediate: Oxalosuccinate -- not a free intermediate,
but an enzyme-bound chemical intermediate.
4. STEP IV: alpha-Ketoglutarate ------> Succinyl-CoA + CO2
1. Oxidative Decarboxylation. Now we have a 4C compound.
2. ENZYME: alpha-Ketoglutarate-Dehydrogenase
Complex.
1. The reaction is similar to pyruvate dehydrogenase
complex.
2. Complex has three enzymes, analogous to E1, E2,
and E3.
3. OXID: alpha-Ketoglutarate ------> Succinyl-CoA
4. Energy in conserved in the COS- (thioester) bond of
-CoA.

3. RED: NAD+ ------> NADH


4. COFACTOR: Coenzyme-A, of course.
5. Product is a thiol-ester -- a high-energy compound.
5. STEP V: Succinyl-CoA ------> Succinate
1. Cleavage of -CoA off of the Succinyl group, breaking a
high-energy bond. This high-energy bond is coupled to
formation of GTP.
2. Substrate-Level Phosphorylation: Direct
phosphorylation of a GDP to produce GTP.
1. GTP comes off here, instead of ATP!
2. Note that this is the only time we get an ATP or GTP
out of the TCA cycle. All other times we get reduced
cofactors (i.e. NADH)
3. ENZYME: Succinate Thiokinase.
4. LABEL-RANDOMIZING STEP: At this point we have a
symmetrical compound. Chirality is lost, and the two
original carbons we invested as Acetyl-CoA will therefore
not be the same ones we get out.
6. STEP VI: Succinate ------> Fumarate
1. (Still 4C): Form a double-bond between the middle-two
carbons. Fumarate is a trans-dicarboxylic acid.
2. ENZYME: Succinate Dehydrogenase.
3. RED: FAD ------> FADH2
4. INHIBITOR -- Malonate is a strong competitive inhibitor of
the enzyme. It blocks the citric acid cycle. It is the cis-acid
of fumarate.
7. STEP VII: Fumarate ------> L-Malate.

1. Hydration Reaction: Add H and OH across the double


bond.
2. STEREOSPECIFIC: Only l-malate is formed, and only lfumarate is a substrate.
3. ENZYME: Fumarase.
8. STEP VIII: L-Malate ------> Oxaloacetate.
1. Oxidation: The -OH group is oxidized to a keto-group.
2. ENZYME: l-Malate Dehydrogenase.
3. Concurrent Reduction: NAD+ ------> NADH
REGULATION OF THE TCA CYCLE:
1. Pyruvate Dehydrogenase: Inhibited by its products -- AcetylCoA and NADH.
1. Part of the NADH influence is substrate deprivation -- that
implies that you have low levels of NAD+ which is required
for the reaction.
2. alpha-Ketoglutarate Dehydrogenase (alpha-Ketoglutarate
------> Succinyl CoA) -- this is the key regulatory step.
1. Ca+ stimulates the step, i.e. Ca+ promotes the TCA cycle.
2. NADH and Succinyl CoA both inhibit the step -- two
products of the reaction.
3. High levels of GTP inhibit the alpha-Ketoglutarate
Dehydrogenase Step.
ANAPLEROTIC REACTIONS: A reaction that provides an essential intermediate
in central metabolism.
1. For example, in order for TCA cycle to function, we must have
Oxaloacetate and Acetyl-CoA. Any source of these
intermediates would then serve as an anaplerotic reaction.

2. Alanine ------> Pyruvate is anaplerotic, because it provides


us with pyruvate, a viable starting point for TCA cycle.
3. Pyruvate ------> Oxaloacetate (in Gluconeogenesis) -- this is
anaplerotic, as it provides us with essential Oxaloacetate.
GLYCOGENESIS / GLYCOGENOLYSIS

STRUCTURE, FUNCTION, AND FLUCTUATION OF GLYCOGEN:


1. Glycogen as an energy source:
1. Fluctuating Levels: Glycogen levels increase and decrease
markedly every day. Generally they increase after a large
meal, and decrease as we expend energy. Glycogen levels
are fairly depleted when we wake up in the morning.
2. Glycogen is a SHORT-TERM energy source. Fat is the longterm energy source.
2. GLYCOGEN STRUCTURE: Glycogen is a highly branched polymer
of pure glucose.
1. Linear Links: alpha-1,4 polyglucose links.
2. Branched Links: alpha-1,6 polyglucose links
3. Reducing Sugar: In a single polymer of glycogen, there
is one sugar that still has a reducing carbon on it at the 1
position. It is the "seed" of the polymer.
4. BRANCHES -- approximately every fourth residue has a
branch.
GLYCOGENOLYSIS: Breakdown of Glycogen to Glucose-1-Phosphate.
1. Glycogen Phosphorylase: Glycogenn ------> Glucose-1Phosphate + Glycogenn-1
1. This is a Phosphorolysis reaction -- not hydrolysis!
2. No ATP is required to break it down. But ATP was required
to build it up, so we're not getting something for nothing.

3. Glycogen Phosphorylase cleaves only the alpha-1,4


linkages -- not the alpha-1,6 linkages.
2. Glycogen phosphorylase takes any glucose chain down to the
last four residues before a branch point. It does not cut beyond
that!
3. Debranching Enzyme: Takes over when within four residues
to a branch point. It has two activities.
1. 4-alpha-D-Glucanotransferase: Transfer 3
residues to another chain on the polymer.
2. Amylo-alpha-1,6-Glucosidase: Hydrolysis of the
final 1,6 bond.
1. This step creates FREE GLUCOSE -- not Glucose-1Phosphate.
2. This is a true hydrolysis reaction -- not
phosphorolysis.
4. Phosphoglucomutase: Glucose-1-Phosphate ------>
Glucose-6-Phosphate.
1. Isomerizes Glucose to a form that can be utilized in
further metabolism.
5. LIVER ONLY: Glucose-6-Phosphatase: Glucose-6Phosphate ------> Glucose ------> Bloodstream.
1. Modulation of blood-glucose level by the liver.
2. Glucagon promotes higher blood glucose, and insulin
promotes lower.
REGULATION OF GLYCOGENOLYSIS: Phosphorylation turns on the system.
1. TWO FORMS OF GLYCOGEN PHOSPHORYLASE: ACTIVATE AND
INACTIVE

1. PHOSPHORYLASE-A: The activated glucose


phosphorylase enzyme. It is phosphorylated to be
activated.
1. Glucose and ATP have inhibitory effects on
Phosphorylase-A. These guys indicate excess
energy, and we don't need to do glycogenolysis in
that case.
2. PHOSPHORYLASE-B: The inactive form of the glucosephosphorylase enzyme. It is dephosphorylated to be
inactivated.
1. AMP has a stimulatory effect on Phosphorylase-B,
and can greatly increase its activity, even without
phosphorylation. AMP has no effect on
phosphorylase-a because its effect is overriden by
the phosphorylation.
1. TWO PHOSPHORYLATING ENZYMES
1. PHOSPHORYLASE KINASE A (ACTIVATOR):
A REGULATORY ENZYME which uses ATP to phosphorylate
Phosphorylase-B, in order to activate it. So, it has an
activating activity.
1. cAMP activates Phosphorylase Kinase A, via a cAMPdependent protein kinase.
2. Ca+2 (from IP3 transduction pathway) increases the
activity of phosphorylase-kinase, as the deltasubunit of the enzyme is acalmodulin which, in this
case, increases the activity of the enzyme when
bound to calcium.
2. PHOSPHOPROTEIN PHOSPHATASE
(DEACTIVATOR): ANOTHER REGULATORY ENZYME which
dephosphorylates Phosphorylase-A, in order to inactivate
it. So, it has an inhibitory activity.

1. INHIBITOR-1A: Inhibits Phosphoprotein


Phosphatase, in order to promote (or sustain)
glycogenolysis.
2. Epinephrine and Glucagon cause the synthesis of
2nd messenger cAMP ------> caMP-dependent Protein
Kinase ------> activates Inhibitor-1A ------> Inhibits
Phosphoprotein Phosphatase ------> PKA remains
phosphorylated ------> glycogen phosphorylase
remains active.
2. THE IMPORTANCE OF CAMP: It not only stimulates the
conversion of phosphorylase-B to phosphorylase-A, but it also
inhibits the conversion of A to B.
1. cAMP turns on Inhibitor 1A in order to inhibit the
Phosphatase ------> Turn on glycogen phosphorylase.
2. Phosphorylates PKA directly ------> generate more
Phosphorylase-A (Active glycogen phosphorylase).
GLYCOGENESIS:
1. YOU MUST START WITH ACTIVATED UDP-GLUCOSE
1. Hexokinase / Glucokinase: Glucose ------> Glucose6-Phosphate
1. ATP required. Glucokinase used in liver, and
hexokinase in muscle.
2. Phosphoglucomutase: Glucose-6-Phosphate ------>
Glucose-1-Phosphate
1. This is the opposite reaction as what occurs at the
end of glycogenolysis.
3. Glucose-1-Phosphate Uridyltransferase: Glucose-1Phosphate ------> UDP-Glucose
1. UTP ------> UMP + PPi. When UMP hooks onto
Glucose-1-Phosphate, it then becomes UDP-Glucose.

2. That means that UDP-Glucose has the UDP attached


to Carbon #1!
3. Pyrophosphatase: Cleaves the high-energy
phosphoric anhydride bond. This drives the
formation of UDP-Glucose and makes its formation
irreversible.
2. GLYCOGEN SYNTHASE: Glycogenn + UDP-Glucose ------>
Glycogenn+1 + UDP
1. Glycogen synthase requires a primer! It will add another
glucose residue to an already-existing chain.
2. Glycogen synthase makes only linear glucose,
connected in alpha-1,4 linkages. A branching enzyme is
required for branching.
3. BRANCHING ENZYME:
1. Once the glycogen chain is about 11 residues long, the
branching enzyme removes about 7 of the residues.
2. It places the 7-residue chain in a 1,6-linkage, at least 4
residues away from any other branching point.
4. GLYCOGENIN: The self-glycosylating protein that serves as the
primer for de novo glycogen synthesis.
1. Usually, no primer is needed:
1. Glycogen Synthase normally has a low Km (i.e. highly
sensitive), given a large starting glycogen molecule.
2. The larger the starting substrate, the lower the
Km for Glycogen Synthase. It won't start with glucose
alone because the Km is way-the-hell too high.
2. GLYCOGENIN ACTIVITY: It has a Serine OH-Group onto
which it adds the first glucose, to start the ball rolling.

1. It takes the first glucose from activated UDPGlucose, releasing UDP (just like the synthase).
2. Glycogenin then dissociates from the growing
glycogen molecule after a few residues have been
put on.
REGULATION OF GLYCOGENESIS: Phosphorylation turns off the system.
1. TWO FORMS OF THE GLYCOGEN SYNTHASE ENZYME: ACTIVE
AND INACTIVE.
1. GLYCOGEN-SYNTHASE A: ACTIVE FORM. It
is dephosphorylated.
2. GLYCOGEN-SYNTHASE B: INACTIVE FORM. It
is phosphorylated.
1. Glucose-6-Phosphate stimulates Glycogensynthase-B to some extent. A surplus of G-6-P
indicates that glycogenesis should take place.
2. INHIBITORY EFFECTS /
PHOSPHORYLATION: Epinephrine and/or Glucagon are
secreted at times of low blood sugar or when we need cellular
energy. They cause a phosphorylation cascade.
1. cAMP, DAG, and Ca+2 (i.e. beta-adrenergic secondary
messengers) all shut off glycogen synthase by promoting
the phosphorylated form of the enzyme. There are two
mechanisms:
1. Direct phosphorylation of glycogen synthase itself,
thereby inactivating it.
2. Phosphorylation of Inhibitor-1 a ------>
phosphorylate (inhibit) the phosphatase ------>
glycogen synthase remains phosphorylated and
hence inactive.

3. STIMULATORY EFFECTS /
DEPHOSPHORYLATION: Glucose stimulates the production of
glycogen.
1. Glucose inhibits Glycogen Phosphorylase-A and the
phosphoprotein phosphatase, as noted above.
1. Phosphorylase-A inactivates Glycogen
Synthase. It is therefore necessary to inactivate
the phosphorylase before we can make glycogen.
2. Glucose therefore stimulates glycogen
synthesis by inhibiting glycogenolysis.
2. Glucose also stimulates Phosphoprotein
Phosphatase.
1. Phosphoprotein Phosphatase itself promotes
glycogen synthesis. It promotes the
dephosphorylated (active) form of Glycogen
Synthase.
INSULIN:
1. It acts by an autophosphorylating tyrosine-kinase receptor
protein.
2. It is generally anabolic: it promotes glycogen synthesis and
inhibits glycogenolysis.
3. It is secreted when glucose levels in the blood are high, and it
promotes uptake of glucose by most cells.
4. It does not affect glucose-uptake by liver cells, because the
liver can uptake as much glucose as it wants by facilitated
transport. Liver absorption of glucose is independent of insulin
levels.
OVERALL ENERGETICS OF GLYCOGEN:

1. GLYCOGENOLYSIS: Every glucose residue released gives us +3


ATP. The end-product of glycogenesis is glucose-6-P, which
can then go through glycolysis.
1. Because the product is glucose-6-phosphate, we don't
need hexokinase in the first step of glycolysis so we save
ourselves 1 extra ATP, giving us a net of +3 ATP (rather
than +2) once the glucose has undergone glycolysis.
2. GLYCOGENESIS: It costs -2 ATP to add on one residue of
Glucose to a glycogen chain. It costs ATP to make glycogen.
1. 1 ATP cost from glucokinase.
2. 1 ATP cost (via UTP) for the glucose-1-phosphate
uridyltransferase
3. NET ENERGY YIELD: ONLY +1 ATP PER GLUCOSE FROM
GLYCOGEN.
4. WHY MAKE GLYCOGEN?
1. We make glycogen in times of plentiful energy. We break
it back down when we need quick energy. Gynogen can
be broken down quickly because it is highly branched.
2. We actually get more ATP (3 -vs- 2) from glycogen then
we do from free glucose, if we disregard the ATP originally
invested.
GLYCOGEN STORAGE DISEASES:
1. Type-I (Von Gierke's Disease): Deficiency of Glucose-6Phosphatase in liver, intestinal mucosa, and kidney
1. Symptoms:
1. Hypoglycemia -- liver can't release glucose!
2. Lactic Acidemia -- Inability to process lactose
normally, as well as increased glycolysis.

3. Increased Uric Acid -- cause not clear.


2. Treatment: Taking small amounts of carbohydrate during
the day to maintain blood glucose can help diminish
symptoms.
2. Type-II (Pompe's Disease): Accumulation of glycogen in
virtually every tissue.
1. Cause: Absence of the alpha-1,4-Glucosidase enzyme,
which normally breaks down polyglucose in lysosomes in
all cells. This leads to obstruction of lysosomal function.
2. Symptom: Ultimately leads to massive cardiomegaly (due
to glycogen in cardiac muscle) and death before 30.
3. Type-III (Cori's Disease): Mild glycogen accumulation, due to
a deficiency in the debranching enzyme.. Essentially,
inefficient utilization of glycogen with a little over storage.
1. Hepatomegaly occurs in early age but then diminishes.
2. Relatively benign condition.
4. Type-V (McArdle's Disease): Absence of Muscle Glycogen
Phosphorylase, so that muscle glycogen stores are
unavailable to exercising muscle.
1. Lab: Increased levels of Muscle CPK (Creatine
Phosphokinase) and myoglobin. These symptoms
generally indicated any muscle disorder.
2. Very serious condition -- defective cardiac muscle.
THE PENTOSE-PHOSPHATE SHUNT

OVERALL PURPOSE:
1. It produces 2NADPH, which is used instead of NADH in most
biosynthetic pathways. This is the primary (perhaps only?)
source of NADPH in metabolism.

1. NADPH is needed in red blood cells as a cofactor


for Glutathione Reductase, which is essential for the
RBC. RBC's can utilize the pentose phosphate shunt to
supply this NADPH.
2. It produces Ribose-5-Phosphate, the essential precursor to
nucleotide synthesis.
PATHWAY:
1. Glucose ------> Glucose-6-Phosphate
1. Catalyzed by Hexokinase; -1ATP
2. Glucose-6-Phosphate ------> 6-Phosphogluconolactone
1. Catalyzed by Glucose-6-Phosphate Dehydrogenase.
This is a REDOX reaction, coupled to NADP.
2. This is an internal-ester -- a lactone ring.
3. Concurrent NADP ------> NADPH Reduction.
3. 6-Phosphogluconolactone ------> 6-Phosphogluconate -a hydrolysis reaction.
1. H2O is added across the lactone to open the ring.
2. Catalyzed by 6-Phosphogluconolactase.
4. 6-Phosphogluconate ------> Ribulose-5-Phosphate -- a
redox reaction
1. Catalyzed by 6-Phosphogluconate Dehydrogenase.
2. It is the oxidation of an alcohol function to a keto group,
and a decarboxylation, leaving a 5-carbon-sugar skeleton.
3. Concurrent reduction of NADP ------> NADPH
5. Ribulose-5-Phosphate ------> Ribose-5-Phosphate

1. This is done via an isomerase.


2. The reactant can also go to Xylulose-5-Phosphate via
an epimerase, but that's not metabolically important.
3. The Ribose-5-Phosphate can then serve as the precursor
for nucleotide synthesis.
6. Through carbon-recombination, Fructose-6-Phosphate is an
ultimate product, which can then be reconverted back to
Glucose-6-Phosphate, which can then start the cycle over
again!
1. Each time the pentose-phosphate pathway recycles as
above, you oxidize the equivalent of one carbon from
glucose. So, if you run the cycle 6 times, you
completely oxidize one glucose.
2. This oxidation of glucose is an alternative to glycolysis
and the TCA cycle, which is not energetically important,
but it does provide essential intermediates -- NADPH and
Ribose-5-Phosphate.
GLUCONEOGENESIS

Q: WHAT CAN BE CONVERTED BACK TO GLUCOSE?


A: Lactate, pyruvate, or any three-carbon equivalent. Acetyl-CoA CANNOT BE
CONVERTED BACK TO GLUCOSE!
LIVER: The main organ that does gluconeogenesis.
MUSCLE: Muscle does gluconeogenesis too if need be, to create glucose-6phosphate. However, it cannot go all the way back to free glucose.
PATHWAY: Starting from Lactate, and going to Glucose
1. Lactate ------> Pyruvate (oxidation)
1. Catalyzed by Lactate Dehydrogenase
2. Concurrent NAD+ ------> NADH (reduction)

2. Pyruvate ------> Oxaloacetate


1. The ultimate goal here is to get back to
Phosphoenolpyruvate, so that we can continue with the
reverse of glycolysis. But since PEP is an irreversible
reaction in glycolysis, we need a workaround.
2. This reaction catalyzed by Pyruvate Carboxylase.
3. ATP is required.
4. This reaction occurs in the mitochondria.
3. Oxaloacetate ------> Phosphoenolpyruvate
1. GTP is consumed. So, in two steps we have gotten to
PEP, at the expense of ATP and GTP, or energetic
equivalent of 2ATP.
2. Catalyzed by Phosphoenolpyruvate Carboxykinase.
3. The malate shuttle allows free passage of this
intermediate to the cytosol, so that gluconeogenesis can
be completed.
4. Phosphoenolpyruvate ------> ------> Glucose-6-Phosphate
1. The rest of the intermediates are the same as they are for
glycolysis.
2. Fructose-1,6-biphosphate ------> Fructose-6-Phosphate
was irreversible in glycolysis, so here it must be catalyzed
by a different enzyme, although we still have the same
intermediates.
ENERGETICS OF GLUCONEOGENESIS: It costs 6 ATP to make one glucose -- 4
ATP + 2 GTP.
LIVER ONLY: Glucose-6-Phosphatase can catalyze Glucose-6-Phosphate ------>
free glucose, which can then be excreted into the blood to raise blood-sugar. This does
not occur in muscles, although muscles undergo gluconeogenesis.

REGULATION OF GLUCONEOGENESIS:
1. Positive Effectors: Generally things that indicate a surplus of
energy.
1. ATP
2. Acetyl-CoA
3. Citrate
2. Negative Effectors: Generally things that indicate a lack of
energy, as well as glycolytic intermediates.
1. AMP
2. Fructose-1,6-biphosphate or Fructose-2,6-biphosphate
3. Inorganic phosphate
NTERCONVERSION OF SUGARS / OLIGOSACCHARIDES / GLYCOPROTEINS

METABOLISM OF GALACTOSE: Galactose (commonly from lactose, or milk


sugar) must be activated to UDP-Galactose before it can be metabolized. Once
activated, it can be freely interconverted with UDP-Glucose for further metabolism.
Starting with free galactose:
1. Galactokinase: Galactose + ATP ------> Galactose-1Phosphate + ADP
2. Glactose-1-Phosphate Uridyltransferase: Galactose-1Phosphate + UDP-Glucose ------> Glucose-1-Phosphate +
UDP-Galactose
3. Galactose-4-Epimerase: UDP-Galactose <====> UDPGlucose
1. This is a two step reaction requiring NAD+ as a cofactor,
although there is no net change in NAD+.
2. Step I: Oxidation of the OH at the position to a carbonyl.

3. Step II: Reduction of the OH at the 4 position, to change


its direction.
GLUCURONIC ACID: Important derivative of glucose, with the C6 carbon
oxidized all the way to a carboxylic acid.
1. It is used in the conjugation of drugs in the liver, to make them
soluble so they can be excreted in the urine.
1. It is a carboxylic acid and thereby negatively charged,
enhancing its ability to solubilize.
2. It is an important component of hetero-polysaccharides.
3. SYNTHESIS = a double-oxidation: C6-OH ------> C6-Aldehyde
------> C6-Acid
1. Concurrent Reduction: 2 NAD+ ------> 2 NADH
GLUCOSAMINE: The acetylated form is an important component of glycoproteins.
Unacetylated, it constitutes the exoskeleton of insects.
1. SYNTHESIS: An amino group is transferred from Glutamine.
Then the product is acetylated.
1. Fructose-6-Phosphate ------> Glucosamine-6Phosphate
1. Concurrent loss of NH3: Glutamine ------>
Glutamate
2. Glucosamine-6-Phosphate ------> NAcetylglucosamine
N-LINKED GLYCOSYLATION: An N-Glycosyl-Link is formed between NAcetylglucosamine and an Asp residue.
1. CONSENSUS SEQUENCE: Asn-X-(Thr or Ser). This is the
consensus sequence found on almost all proteins where NLinked Glycosylationmight occur.

1. X can be any residue except Proline.


2. There is a CORE of two N-Acetylglucosamines and 2 mannoses
that are hooked to the Asn residue.
3. 20-50 more sugars are attached. Most sugars are found in the
chains, but typically not N-Acetylgalactosamine.
4. N-Linked sugars are stable in alkaline environment.
O-LINKED GLYCOSYLATION: N-Acetylgalactosamine is the more common
sugar. It is attached via a hydroxy group to a Ser or Thr residue.
1. O-Linked sugars are unstable in alkaline environment.
2. The sugar chain can be hundreds of sugars long.
3. O-Links often contain N-Acetyl-Neuraminic
Acid and Fucose, but never Glucuronic Acid.
4. Linked by 1,2 and 1,3 linkages to each other.
HYDROXYLYSINE: Found in collagens. Glucose residues, in groups of only 2 or 3,
are commonly glycosylated to Hydroxylysine residues on collagen-chains.
DOLICHOL PHOSPHATE: Aids in N-Linked glycosylation in the Golgi
Membrane (or associated with it).
1. Dolichol Phosphate hooks up with charged UDP-NAcetylglucosamine, to form a sugar with a long fatty tail.
Then another residue of the same thing comes.
2. Then multiple mannoses are added, charged in the form
of GDP-Mannose.
3. In the end, we have a sugar core of a series of mannoses, and
two N-Acetylglucosamines.
4. In this form, the Dolichol Pyrophosphate complex transfers the
sugar groups to a suitable protein residue.

HETEROPOLYSACCHARIDES: Found in the extracellular matrix, synovial fluid,


lubricants, etc.
1. CHONDROITIN SULFATE: A polymer of glucuronic acid and NAcetylgalactosamine sulfate.
1. Hydroxy group at C4 of the N-Acetylgalactosamine is
sulfated.
2. It is a highly charged polymer, found in ground
substance, having a high water of hydration.
2. HEPARIN: Heteropolysaccharide -- natural anti-coagulant.
1. A disaccharide of glucuronic acid and NAcetylglucosamine-6-Phosphate.
2. It is extremely negatively charged with one carboxylate
and three sulfates.
3. Produced in high quantities in the lungs.