The PPP & Fructose, Galactose

and GAGs Metabolism

Prof. LOZANO-DE LA ROSA, Cuauhtémoc

Department of Biochemistry and Molecular Biology
International Program, School of Medicine
[email protected]
 The Pentose Phosphate Pathway
• The Pentose Phosphate Pathway (PPP) is an anabolic pathway
that utilizes glucose to generate pentose sugars and reducing
equivalents. The primary functions of this pathway are:
1. To generate the NADPH required for the synthesis of Fatty Acids,
Cholesterol, steroid hormones and ketone bodies. The reduction

of oxidized Glutathione (GS-SG), and the Cytochrome P450.

2. To provide the cell with Ribose-5-P (R-5-P) for the synthesis of
nucleotides for the synthesis of nucleic acids and coenzymes.
3. The PPP can operate to metabolize dietary pentose sugars
derived from the digestion
of nucleic acids as well as
to rearrange the carbon
skeletons of dietary
carbohydrates into
glycolytic/gluconeogenic
intermediates.
 The Pentose Phosphate Pathway

Synthesis/function of:
*Cholesterol
*Steroid Hormones
*Fatty Acids
*Ketone Bodies
*Reduction of oxidized
Glutathione (GS-SG)
*Mixed Function Oxidases
*Cytochrome P450
 The Pentose Phosphate Pathway

To Nucleotide
synthesis used
for Nucleic acids
and Coenzymes
TPP structure

(TPP)

Glycolysis and
Gluconeogenesis
intermediates
TPP
 The PPP Reactions
• No ATP is directly consumed or produced in the cycle
• Carbon 1’ of glucose 6-phosphate (G-6-P) is released as CO 2, while
2 NADPH are produced for each G-6-P entering the oxidative part
of pathway
• Rate and direction of reversible reactions are determined by
availability of intermediates

A. Glucose 6-P becomes 6-phosphogluconolactone, while NADP + is

reduced to NADPH(H+)
Enzymes: Glucose 6-phosphate Dehydrogenase &
6-Phosphogluconolactone hydrolase
B. 6-phosphogluconolactone is hydrolyzed to 6-phosphogluconate
Enzyme: Gluconolactonase
 The PPP Reactions
C. 6-phosphogluconate is then oxidatively decarboxylated, releasing
CO2, a second NADP+ is reduced, producing NADPH(H+), and the
remaining carbons form Ribulose-5-phosphate
Enzyme: 6-Phosphogluconate Dehydrogenase
D. Ribulose-5-P isomerizes to Ribose-5-P or epimerizes to
Xylulose 5-P
Enzymes: Ribose 5-Phosphate Isomerase &
Phosphopentose Epimerase
E. Ribose 5-Phosphate and Xylulose 5-P then undergo reactions to
form Fructose 6-P and Glyceraldehyde-3-P
Enzymes: Transketolase (coenzyme: Thiamine
Phyrophosphate, TPP) & Transaldolase
Transketolase ∆ 2C units & Transaldolase ∆ 3C units
 Synthesis of dNDPs
• The conversion of ribonucleotides to 2’-deoxyribonucleotides
(through the action of Ribonucleotide reductase) requires NADPH
as the electron source, therefore, any rapidly proliferating cell needs
large quantities of NADPH.
 Fatty Acid Synthesis
• The reactions of fatty acid biosynthesis and steroid biosynthesis
utilize large amounts of NADPH. Hepatocytes, adipocytes, adrenal
cortex, testis and lactating mammary gland have high levels of the
PPP enzymes. In fact 30% of the oxidation of glucose in the liver
occurs via the PPP
• NADPH serves as electron donor in the two reduction reactions.
 The Oxygen Burst
• Although the PPP operates in all cells,
the highest levels of PPP enzymes
(in particular Glucose 6-phosphate
Dehydrogenase) are found in neutrophils
and macrophages. These leukocytes are
the phagocytic cells of the immune system
and they utilize NADPH to generate
superoxide radicals (O2.-) from molecular
oxygen in a reaction catalyzed by the
enzyme NADPH oxidase.
The superoxide anion, in turn, serves to
generate other reactive oxygen species
(ROS) that kill the phagocytized microorganisms.
Following exposure to bacteria and other foreign
substances there is a dramatic increase in O2 consumption by
phagocytes. This phenomenon is referred to as the oxygen burst.
The Oxygen Burst
 Chronic Granulomatous Disease
(CGD)
• CGD is usually inherited in an X-linked recessive fashion. Most
patients (90%) are males and have defects on the gene coding for
gp91phox, p22phox, p47phox or p67phox. The autosomal
recessive (AR) forms may be associated with milder disease.

Pathophysiology
• CGD is a disorder resulting from the inability of phagocytes to kill
microbes they have ingested. The normal functioning reaction is
mediated by the phagocyte NADPH Oxidase also called Phagocyte
Oxidase (phox). When this is non functioning there is a defect in the
killing mechanism caused by any of several defects in the NADPH
oxidase enzyme complex which generates the microbicidal
respiratory burst
• Infections are caused mostly by catalase positive organisms, such as
Aspergillus spp. or Staphylococcus spp., and are often life-
threatening.
 Chronic Granulomatous Disease
• In phagocytes, the NADPH oxidase complex is activated to produce
Superoxide anion (O2.-) and other secondarily derived ROS
(reactive oxygen species), which promote killing of invading micro-
organisms.
 Chronic Granulomatous Disease
• Patients with CGD experience severe recurrent bacterial and fungal
infections and often develop granulomas, which may form initially by
accumulation of phagocytes containing ingested bacteria. The
accumulation of phagocytes occurs because they are unable to kill
ingested pathogens and because apoptosis/turnover is abnormal,
both of which are due to the defect in NADPH oxidase function.
 G-6-P Dehydrogenase Def.
• Glucose 6-phosphate Dehydrogenase
(G6PD) is a house-keeping enzyme
critical in the redox metabolism of all
aerobic cells. G6PD deficiency has been
a prototype of hemolytic anemias due to
enzymopathy, i.e. to a primary
abnormality of a red blood cell enzyme.
G6PD deficiency is also a prime example
of a hemolytic anemia due to an
interaction between an intracorpuscular
cause and an extracorpuscular cause,
because in the majority of cases
hemolysis is triggered by an exogenous
agent.
• Glucose-6-P dehydrogenase deficiency is
an X-linked inherited disease
 G-6-P Dehydrogenase Def.

• At least 400 million people carry a G6PD deficiency gene. Areas of high
prevalence are Africa, Southern Europe, the Middle East, South-East Asia
and Oceania. In the Americas and in parts of Northern Europe. G6PD
deficiency is also quite prevalent as a result of migrations in relatively
recent historical times
 G-6-P Dehydrogenase Def.
• Most individuals who have inherited a G6P dehydrogenase mutation do not show clinical manifestations.
However, some patients with G6PDH deficiency develop hemolytic anemia because of the following
reasons:

1. Treated with an oxidant drug: Antibiotics:

(Sulfamethoxazole), Antimalarials:
(Primaquine, Pamaquine but not Quinine).
The ring of Pamaquine, has a pyridinium ring whose
structure resembles the pyridinium ring of NADP.
Antipyretics: (Acetanilid but not acetaminophen)

2. Ingestion of fava beans (vicia faba) Some forms of Pamaquine

G6PD deficiency are susceptible to fava beans.
Favism is not observed in all individuals with G6PD
deficiency. However, all patients with favism have
G6PD deficiency

3. Contract a severe infection. Most common

precipitating factor of hemolysis in G6PD deficiency.
Inflammatory response results in generation of free
radicals in macrophages. Free radicals enter red blood
cells, causing oxidative damage
Faba beans glycosides

Vicine
 G-6-P Dehydrogenase Def.
Typically, a hemolytic attack starts with malaise, weakness, and
abdominal or lumbar pain. The onset can be extremely abrupt,
especially with favism in children.
After an interval of several hours to 2-3 days, hemoglobin spills
over into the urine, resulting in hemoglobinuria and and dark-
colored urine.
Bilirubin, a product of heme catabolism also accumulates in blood
leading to jaundice (yellowish discoloration of the skin and
sclera).
There are over 400 different mutations of the G-6-PD gene but some
of these mutations cause clinical symptoms because an unstable
enzyme that has a shorter half in the RBC or an enzyme
that is unusually sensitive to inhibition by low levels of NADPH,
infections or the purine glycosides of fava beans. In all cases, the
cell’s ability to recycle: G-S-S-G + NADPH  2 G-SH + NADP+
is impaired, and drug induced oxidative stress may lead to
hemolysis.
 Glutathione (G-SH)
• Maintenance of the integrity of the RBC membrane depends on its
ability to generate ATP and NADH from glycolysis. NADPH is
generated by the PPP and utilized for the reduction of oxidized
glutathione: G-S-S-G + NADPH  2 G-SH + NADP+.
• Glutathione is a tripeptide that includes a g-glu-cys-gly residues. Its
functional group is the cysteine thiol (-SH). Glutathione is
necessary for the removal of H2O2 and lipid peroxides generated by
reactive oxygen species (ROS) in the oxygen-rich environment
within RBCs.
 NADPH and Oxidative Stress
Glutathione Peroxidase catalyzes degradation of peroxides by
reduction, as two glutathione molecules are oxidized to a disulfide.
2 G-SH + R-OOH  G-S-S-G + ROH + H2O
Regeneration of reduced glutathione requires NADPH, produced
within erythrocytes in the PPP. Glutathione Reductase catalyzes:
G-S-S-G + NADPH(H+) 2 GSH + NADP+
 G-6-P Dehydrogenase Def.

Glutathione also helps

mantain the reduced
states of sulfhydryl (-SH)
groups in proteins,
including hemoglobin.
Oxidation of those
sulfhydryl froups leads to
the formation of denatured
hemoglobin that form
insoluble masses called
Heinz bodies (HzB) that
attach to the red blood cell
membranes and
hemolysis occurs.
 G-6-P Dehydrogenase Def.
• The negative effect of G-6-PD
deficiency has been balanced
by an increased resistance to
malaria shown by
female carriers
of the mutation.
• G6PD is one of the best
characterized enzyme protein
polymorphisms in the human.
species. Clinical field studies
and
in vitro experiments
strongly support the view that
G6PD deficiency
has been
selected by Plasmodium
falciparum malaria, by virtue of
the fact that it confers a
relative resistance
to heterozygotes, and
perhaps to hemizygotes as well, against this highly
lethal infection. Different G6PD variants underlie
G6PD deficiency in different parts of the world.
 Metabolism of other Sugars
Ga(1-1)aG Galb(1-4)bGlu

Glua(1-2)bFru
 Sucrose Digestion
• Sucrase-Isomaltase (SI)
(EC 3.2.1.10), is one of four integral
glycoproteins expressed in the brush
border membrane of the small
intestine. Sucrase-isomaltase is a
hydrolase enzyme responsible for
catalyzing the hydrolysis of dietary
sucrose and starch into glucose
and fructose for transport into the
blood stream.
 Fructose Absorption
• A deficiency of Sucrase-Isomaltase, can inhibit sucrose digestion
and absorption. This allows sucrose to pass undigested through the
intestines and serve as fuel for the naturally occurring bacteria. This
bacterial metabolism results in excessive gas, cramping, bloating,
abdominal pain, constipation and diarrhea. The lack of glucose
absorption decreases energy production and impairs many
biochemical processes, which can affect growth and development.
 Fructose Metabolism
• Muscle, which contains two types of Hexokinase (type I and type II),
can phosphorylate fructose to F6P which is a direct glycolytic
intermediate. However, the affinity of hexokinase for fructose is
substantially less than that of fructokinase.
• In the liver, which contains mostly Glucokinase (Hexokinase type IV),
which is specific for glucose as its' substrate, there is the requirement
for KHK (Ketohexokinase: Fructokinase) to utilize fructose in
glycolysis. Hepatic KHK-C phosphorylates fructose on C–1 yielding
Fructose-1-phosphate (F1P). In liver the form of Aldolase that
predominates (Aldolase B) can utilize both F-1,6-BP and F1P as
substrates. Therefore, when presented with F-1P the enzyme generates
DHAP and glyceraldehyde. The DHAP is converted, by triose
phosphate isomerase, to GA3P and enters glycolysis. The
glyceraldehyde can be phosphorylated to GA3P by Glyceraldehyde
kinase or converted to DHAP through the concerted actions of Alcohol
dehydrogenase, Glycerol kinase and Glycerol phosphate
dehydrogenase.
 Fructose Metabolism

• Enzymes of fructose metabolism: (1) phosphohexose isomerase; (2) hexokinase;

(3) sorbitol dehydrogenase; (4) fructokinase; (5) aldolase-B; (6) alcohol
dehydrogenase; (7) aldehyde dehydrogenase; (8) triose kinase; (9) triosephosphate
isomerase; (10) glycerol 3-phosphate dehydrogenase; (11) glycerol kinase; (12)
glycerate kinase; (13) glyceraldehyde 3-phosphate dehydrogenase; (14) fructose
diphosphatase; (15) phosphoglycerate kinase; (16) phosphoglyceromutase
 Hepatic Fructose
Metabolism
In most cells, fructose is
metabolized just the way glucose
is. Liver cells don't work this way.
Glucokinase cannot work on
fructose. Fructokinase, therefore,
is the enzyme that catalyzes the
phosphorylation of fructose
producing Fructose-1-P
Eventually, it is converted into one
of the glycolytic intermediates,
and can be properly run through
mitochondria after that. BUT, this
bypasses the enzyme that is
subject to feedback regulation.
Therefore, while glucose is
readily stored as glycogen,
fructose is not. Fructose is
processed into pyruvate, and
transformed to Acetyl-SCoA which
is used in fatty acid synthesis and
triglycerides synthesis
 Non-liver cells Fructose
Metabolism

In most cells, fructose is

metabolized just the way
glucose is. First the enzyme
Hexokinase, transfers a
phosphate from ATP to the
sugar. The enzyme produce
Glucose-6-P (from glucose)
and Fructose-6-P (from
fructose). The second enzyme
of glucose metabolism converts
G-6-P to Fructose-6-P.
Consequently, both sugars are
metabolized the same way.
The thumbnail on the right
highlights the processing of
fructose by this mechanism.
Fructose Metabolism
 Essential Fructosuria
• Deficiency of Fructokinase (Ketohehokinase, KHK)
• Mutations in the KHK gene, located on chromosome 2p23.3-23.2
are responsible
• Fructose is either excreted unchanged in the urine or metabolized
to Fructose-1-phosphate by alternate pathways in the body, most
commonly by Hexokinase in adipose tissue and muscle.
• Autosomal recessive
• 1:130,000 births
• Benign metabolic defect
• There is no abnormality other than
excretion of fructose in urine.
The excretion of fructose in the urine is not
constant, it depends largely on dietary
intake.
• Diagnosis is typically made after a positive
• test for reducing substances in the urine.
 Hereditary Fructose Intolerance
• Autosomal recessive fructose metabolism disorder
• Defective Fructose-1-P Aldolase (Aldolase B) (EC 2.1.2.13)
• Accumulation of Fructose-1-P that inhibits Glycogenolysis and
Gluconeonesis by inhibition of Glycogen Phosphorylase and F-1,6-BP
Aldolase A impairing condensation of GA-3’P and DHAP to form F-1,6-BP
that then goes gluconeogenesis, causing severe hypoglycemia following
ingestion of fructose.
• Hypoglycemia causes sweating, convulsions ans absent reflexes.
• Pi is sequestred as F-1-P causing low levels of Pi and ATP
• Uncontrolled consumption of ATP leads to intracellular Pi depletion and
activation of AMP deaminase leading to increased production of uric
acid
• Lethargy, lose weight and fructosuria
• Prolonged fructose ingestion in infants leads ultimately to hepatic damage,
jaundice and death.
• Patients develop a strong distaste for sweet food, and avoid a chronic
course of the disease by remaining on a fructose- and sucrose-free diet.
Fructose Metabolism

AD
 Galactose Metabolism
• Galactose, is metabolized from the milk disaccharide, Lactose, after
digestion by Lactase (b-Galactosidase, EC 3.2.1.23), an enzyme of
the brush border of the intestinal epitelium (microvilli).

• At weaning, when diet is changed from a milk-based to a mixed adult

diet, the small intestine undergoes functional maturation. Lactase
activity decreases and its longitudinal distribution is modified, while
the activity of other enzymes, such as sucrase-isomaltase (EC
3.2.1.10), increases.

•
 Galactose Metabolism
• Galactose metabolism occurs through a series of steps that is referred to
as the Leloir pathway.

• First the galactose is phosphorylated by Galactokinase to yield Galactose-

1-phosphate (Gal-1-P). Galactokinase is encoded by the GALK1 gene
located on chromosome 17q25.1. Epimerization of galactose-1-phosphate
to G1P requires the transfer of UDP from uridine diphosphoglucose (UDP-
glucose) catalyzed by Galactose-1-P Uridyltransferase (GALT). The GALT
gene is located on chromosome 9p13.3. The GALT catalyzed reaction
generates UDP-Galactose and G1P.

• The UDP-galactose is epimerized to UDP-glucose by UDP-Galactose-4’

epimerase (GALE). The UDP portion is exchanged for phosphate
generating G1P which then is converted to G6P by Phosphoglucose
mutase. The GALE gene is located on chromosome 1p36.11. GALE
catalyzes two distinct but analogous epimerization reactions, the
epimerization of UDP-galactose to UDP glucose and the epimerization of
UDP-N-acetylgalactosamine to UDP-N-acetylglucosamine.
Galactose Metabolism

Waldenase (EC5.1.3.2)
Galactose Metabolism
 Galactokinase Deficiency
Galactokinase deficiency
(Galactosemia type II)

• Galactosemia and galactosuria

• Symptoms are milder because Gal-1-P
is not formed and hence no toxic effects
of this compound are manifested,
• urine may be positive for reducing
substances.
• Elevated Galactitol may cause
development of cataracts usually in the
neonatal period, but occasionally delayed
until adulthood.
• Frequency: 1 in 40,000 births
• Treatment: Restriction of galactose
Classic Galactosemia


 Classic Galactosemia
• Deficiency of Gal-1-P Uridyl Transferase (Galactosemia Type I).
• AR inheritance
• Frequency: 1 in 35,000 births
• Symptoms of galactosemia, which usually begins within a few days of
ingesting breast milk or lactose-containing formulas, are:
• Gastrointestinal findings: Vomiting and Diarrhea
• Feeding problems: baby refuses to eat formula containing milk;
• If the diagnosis of galactosemia is not made in the neonatal period, failure
to thrive, chronic vomiting, hepatic cirrhosis, and mental retardation may
develop in infants who survive.
• Hyperammonemia
• Hepatocellular damage and especially jaundice of intrinsic liver disease:
Bilirubin uptake is less and bilirubin conjugation decreases, so
unconjugated bilirubin increases resulting in jaundice
• Gal-1-P accumulates in the liver. This will inhibit Galactokinase and
Glycogen phosphorylase resulting in hypoglycemia
 Classic Galactosemia (cont.)

• Neurological findings: Severe mental retardation, irritability and

seizures, lethargy
• Free galactose accumulates leading to galactosemia and
galactosuria
• Gal-1-P may get deposited in renal tubules causing damage and
generalized aminoaciduria (Renal Fanconi's syndrome)
• Escherichia coli sepsis has been described in many patients with
galactosemia.
• Diagnosis: clinical manifestations including congenital cataract,
galactosemia and galactosuria. Collection of fetal cells by
amniocentesis
• Treatment: Lactose-free diet for affected infants. After four years
the diet may be withdrawn when Gal-1-P pyrophosphorylase (*)
becomes active. Gal-1-P + UTP (*)  UDP-Galactose + 2Pi
 The Polyol Pathway
• In galactosemic patients, galactose becomes the substrate for
the polyol pathway: Aldose reductase reduces galactose to
galactitol. Galactitol, accumulates in body tissues and is excreted
in the urine of galactosemic patients. Accumulation of galactitol has
been attributed to galactosemic cataract, and high concentrations
of galactitol have been found in people with classic galactosemia.
Galactose concentration must be fairly high before the enzyme,
aldose reductase, will convert significant amounts of the sugar to
its galactitol form.
 GALT Mutations
• The gene for Galactose-1-phosphate Uridylyltransferase ( GALT gene )
is located on the short arm of chromosome 9, in the region 9p13.

• There are several clinical variants due to genetic mutations in Gal-1-PUT

that alter, but do not eliminate, enzyme activity. The most common of these
are the Duarte (Asn314Asp) and Los Angeles variants. Patients with these
variants are usually clinically asymptomatic; however the reduced enzyme
activity will be detected by newborn screening. Further testing is required.

• Most changes in the GALT gene alter a single amino acid. The most
common GALT mutation in Caucasian Europeans and North Americans is
Gln188Arg. Another mutation occurs almost exclusively in people of African
descent. Is Ser135Leu.

• A particular GALT mutation called the Duarte variant results in a form of

galactosemia with less serious complications than the classic type.
The signs and symptoms associated with this variant tend to be milder
because the enzyme retains 5 percent to 20 percent of its normal activity.
 Prenatal Testing
• Analysis of GALT enzyme activity and molecular diagnosis rely
on cells obtained by chorionic villus sampling (CVS) at
approximately 10 to 12 weeks' gestation or amniocentesis usually
performed at approximately 15 to 18 weeks' gestation. When a fetus
has GALT deficiency, amniotic fluid concentration of Galactitol is
elevated in the late third trimester.
Classic Galactosemia
 Synthesis of Lactose

• Lactose is the “milk sugar” produced by the

mamary glands of most mammals.
• Lactose is synthesized in the Golgi by
Lactose Synthase (UDP-Galactose:Glucose
Galactosyltransferase), which transfers
galactose from UDP-galactose to glucose,
releasing UDP.
• The enzyme is a dimer composed by two
proteins: protein A and protein B.
• Protein A is a b-d-Galacosyltransferase and is
found in several tissues to synthesize
structurally important N-linked glycoproteins.
• Protein B is found only in lactating mammary
glands. It’s a a-Lactalbumin, and its synthesis
is stimulated by the peptide hormone
Prolactine
 Mannose Metabolism
• The digestion of many polysaccharides and glycoproteins yields
mannose which is phosphorylated by Hexokinase to generate
Mannose-6-phosphate. Mannose-6-phosphate is converted to
Fructose-6-phosphate, by the enzyme Phosphomannose
isomerase, and then enters the glycolytic pathway or is converted
to glucose-6-phosphate by the gluconeogenic pathway of
hepatocytes.
 Glycomics

• The nine common

sugars found in
mammalian cells can be
combined in a myriad
number of ways to form
complex carbohydrate
structures (glycans).
The glycan repertoire
(glycome) of a given
cell or organism is thus
many orders of
magnitude more
complex than the
genome or the
proteome.
Glycoproteins
Glycosaminoglycans (GAGs)
 Glycosaminoglycans
• Glycosaminoglycans (GAGs) or Mucopolysaccharides are
unbranched polysaccharides composed of repeating disaccharide
structures that are typically sulfated at specific sites on the
backbone.
 Glycosaminoglycans
• Glycosaminoglycans, GAGs or
Mucopolysaccharides, are polymers of of
anionic heteropolysaccharides chains,
associated with a small amount of
protein.
• GAGs bind large amounts of water forming
the gel-like matrix that forms the basis of the body ground substance.
• GAGs stabilize and support cellular and fibrous components of tissue while
helping maintain the water and salt balance of the body. Connective tissue in
skin, tendons, cartilage ligaments and the bone matrix, consist of
insoluble protein fibers distributed in the ground substance, whereas tendon
is composed primarily of fiber. Synovial fluid serves as a lubricant in joints,
tendon sheets, and bursa.
 Glycosaminoglycans
• Glycosaminoglycans (GAGs) are unbranched heteropolysaccharides
composed of repeating disaccharide structures that are typically sulfated at
specific sites on the backbone.
 GAGs Functions
• Some specific functions of GAGs are:

• Hyaluronic acid: Cell migration in embryogenesis, Morphogenesis

and wound healing
• Chondroitin Sulfate: Formation of bone, cartilage and cornea
• Keratan Sulfate: Transparency of cornea
• Dermatan Sulfate: Transparency of cornea. Binds chylomicrons to
endothelial cells from capillary walls for degradation by Lipoprotein
Lipase
• Heparin: Anticoagulant (it binds Antithrombin III). It anchors
Lipoprtein Lipase to epithelial cells of capillary walls
• Heparan Sulphate: Conmponent of skin fibroblasts and aortic wall;
commonly found on cell surfaces
 GAGs Functions
• Heparin binds to
antithrombin III
(AT III), altering the
conformation and
enhancing the
activity of this major
protease inhibitor.
• Heparan Sulfate
binds Lipoprotein
Lipase (LPL) that is
activated by ApoC-II
present in
chylomicrons and
VLDL for lipolysis of
triglycerides.
 Synthesis of GAGs
The core protein is synthesized at the rough endoplasmic reticulum.
The protein is then glycosylated by specific membrane-bound
Glycosyltransferases located in the Golgi complex.
Synthesis of L-iduronic acid occurs after D-glucuronic acid has been
incorporated into the carbohydrate chain. Uronosyl-5-epimerase
causes epimerization of the D- to the L-sugar.
Addition of sulfate groups occurs after the monosaccharide to be
sulfated has been incorporated into the growing carbohydrated
chain. 3’Phosphoadenosyl-5’-phosphosulfate (PAPS) is the
source of sulfate.
The polysaccharide chains are
elongated by a sequential
addition of alternating acidic
and amino sugars, donated by
their UDP-derivatives

PAPS
Synthesis of Chondroitin Sulfate
 Synthesis of GAGs
• The GAG, on a serine residue of the core protein, is synthesized in a
pathway that begins in the endoplasmic reticulum and concludes in the
Golgi apparatus. It requires several specific Glycosyltransferases.
Synthesis of GAGs
 Synthesis of N-Acetyl Aminosugars
• The monosaccharide Fructose 6-phosphate is the precursor of
GlcNAc, GalNAc, and the Sialic acids, including N-Acetylneuraminic
acid (NANA). In each of these sugars, a hydroxyl group of the
precursor is replaced by an amino group donated by glutamine
 Glucuronic Acid
• Glucuronic acid can be obtained in small
amounts from the diet and from the intracellular
lysosomal degradation of GAGs, or via the uronic
acid pathway. The end product of glucuronic acid
metabolism in humans is Xylulose-5-phosphate,
which can enter the hexose monophosphate
pathway and produce the glycolytic intermediates
GA3’-P and F6-P. The active form of glucuronic
acid is UDP-glucuronic acid, which is produced
by oxidation of UDP-glucose
 Mucopolysaccharidoses (MPS)
• The mucopopysaccharidoses are hereditary disorders caused by
a deficiency of any one of the lysosomal hydrolases normally
involved in the degradation of Heparan sulfate (HS) and/or
Dermatan sulfate (DS). This results in the presence of
oligosaccharides in the urine, because incomplete lysosomal
degradation of GAGs. These fragments are used to diagnose the
specific mucopolysacharidosis, namely by identifying the sugar
residue present on the nonreducing end of the oligosaccharide.
That residue would have been the substrate for the defective
enzyme
• The mucopolysaccharidoses are characterized by accumulation of
GAGs in various tissues, causing varied symptoms such as
skeletal and extracellular matrix deformities, and mental
retardation.
• All muchopolysaccharidoses are autosomal and recessively (AR)
inherited, except Hunter’s syndrome, which is X-linked
Degradation of DS and HS
Degradation of KS
Degradation of GAGs
 Hurler Syndrome (MPS I)
• Hurler Syndrome, MPS I-H
(Gargoylism)
Most severe form of MPS I
a-L-Iduronidase deficiency
Mental retardation, Skeletal deformities,
Corneal clouding, dwarfing, coarse
facial features, upper airway obstruction,
Degradation of DS and HS are defective
and its deposition in coronary artery
leads to ischemia and early death.
Treated by bone marrow transplantation,

preferably before 18 moths.

 Hunter Syndrome (MPS II)

• Iduronate sulfatase
deficiency
• Degradation of DS and HS
are defective. DS and HS in
urine
• X-linked deficiency
• No corneal clouding but
skeletal deformities and
mental retardation is mild
to severe.
• Deafness.
 Sanfilippo’s Syndrome (MPS III)
Four different types are reported. Every enzyme
catalyzes a step necessary for removal of N-
sulfated or N-Acetylated glucosamine residues
from heparan sulfate

• Type A: Heparan sulfatase deficiency

• Type B: N-Acetylglucosaminidase deficiency
• Type C: Glucosamine-N-Acetyltransferase
deficiency
• Type D: N-Acetylglucosamine-6-sulfatase
deficiency

• Severe nervous system disorders. Mental

retardation. Skeletal deformity. Corneal
clouding. Heparan sulfate in urine
 Morqio’s Syndrome (MPS IV)
Two different types are reported
1. Galactosamine sulfatase deficiency
2. b-D-Galactosidase deficiency
• Mental retardation, skeletal deformities, epiphysial dysplasia,
corneal clouding, abnormal heart development, hypermobile joints,
widely spaced teeth, dwarfism, bell-shaped chest (flared ribs)
• Keratan sulfate and Choindroitin sulfate in urine
 Scheie’s Syndrome (MPS V)
• L-Iduronidase
deficiency
• No mental retardation,
mild skeletal changes,
corneal clouding,
cardiac valve
abnormalities, joint
contractures, Carpal
tunnel syndrome,
hernia, coarse facial
features and
hepatomegaly
• Dermatan sulfate in
urine
 Maroteaux-Lamy’s Syndrome (MPS VI)
• N-Acetyl-b-D-Galactosamine-4-
Sulfatase Deficiency (Aryl
sulfatase B Deficiency)
• Patients share many of the
physical symptoms found in
Hurler syndrome
• No mental retardation, skeletal
deformities particularly in the
pelvic region, shortened trunk
and crouched stance, protruding
abdomen and forward-curving
spine, corneal clouding,
deafness, thickening of the dura
• Dermatan sulfate in urine
 Sly’s Syndrome (MPS VII)
• b-Glucuronidase deficiency
• The symptoms of Sly syndrome are similar to those of Hurler
syndrome (MPS I).
• Mental retardation, skeletal deformities: pectus carinatum or
excavatum, kyphosis or scoliosis, short stature, corneal clouding,
Hepatosplenomegaly
• Impaired degradation of Dermatan sulfate and Heparan sulfate
As usual…..