Carbohydrates Metabolism

GLYCOLYSIS & THE OXIDATION OF

PYRUVATE
 Introduction to metabolism
Definition of metabolism.1
.Types of metabolism.2
DIGESTION OF CARBOHYDRATES
.Abnormalities in disaccharides digestion
Absorption of monosaccharides by
. intestinal mucosal cells
1.Galactose and Glucose are transported into
the mucosal cells by an active, energy-
requiring process that involves a specific
transport protein and requires a concurrent
uptake of sodium ions.
2. Fructose uptake requires a sodium-indepen-
dent monosaccharide transporter for its
absorption.
.Transport of Glucose into the cells
A.Na+-independent facilitated diffusion
transport .
At least 14 types-GLUT-1 to GLUT-14.
1.GLUT-1 is abundant in erythrocytes and brain, but is low in
adult muscle.

2.GLUT-2, which is found in the liver, kidney, and beta

cells of the pancreas, can either transport glucose into these
cells when blood glucose levels are high, or transport glucose
from the cells to the blood when blood glucose levels are low
(for example, during fasting).
•
3.GLUT-3 is the primary glucose transporter in
neurons.
4.GLUT-4 is abundant in adipose tissue and
skeletal muscle. The number of GLUT-4
transporters active in these tissues is
increased by insulin.
5.GLUT-5 is unusual in that it is the primary
transporter for fructose (instead of glucose) in
the small intestine and the testes.

6.GLUT-7, which is expressed in the liver and other

gluconeogenic tissues, mediates glucose flux
across the endoplasmic reticular membrane.
B.Na+-monosaccharide cotransporter system.
Energy-requiring process that transports glucose "against"
a concentration gradient from low glucose concentrations
outside the cell to higher concentrations within the cell .

Movement of glucose is coupled to the concentration

gradient of which is transported into the cell at the same
time.
This type of transport occurs in the epithelial cells of the
intestine, renal tubules, and choroid plexus.
GLYCOLYSIS & THE OXIDATION OF PYRUVATE

Definition.
Types.
Reactions.
Regulation.
Clinical importance.
 Glycolysis
• Glycolysis is a cytoplasmic pathway that
converts glucose into two pyruvates,
releasing a modest amount of energy
captured in two substrate-level
phosphorylations and one oxidation
reaction.
• Glycolysis also provides intermediates for
other pathways. In the liver, glycolysis is
part of the process by which excess glucose
is converted to fatty acids for storage.
 Hexokinase and glucokinase:
• Glucose entering the cell is trapped by
phosphorylation using ATP.
• Hexokinase is widely distributed in tissues
whereas glucokinase is only found in
hepatocytes and pancreatic b-islet cells.
• Phosphofructokinases (PFK-1 and PFK-2): PFK-1 is the
rate-limiting enzyme and main control point in glycolysis. In
this reaction, fructose 6-phosphate is phosphorylated to
fructose 1,6-bisphosphate using ATP.
• Glyceraldehyde 3-phosphate dehydrogenase—catalyzes an
oxidation and addition of inorganic phosphate (P i) to its
substrate.
• This results in the production of a high-energy intermediate
1,3-bisphosphoglycerate and the reduction of NAD to
NADH.
• If glycolysis is aerobic, the NADH can be reoxidized
(indirectly) by the mitochondrial electron transport chain,
providing energy for ATP synthesis by oxidative
phosphorylation.
• 3-Phosphoglycerate kinase—transfers the high-
energy phosphate from 1,3-bisphosphoglycerate
to ADP, forming ATP and 3-phosphoglycerate.
• This type of reaction in which ADP is directly
phosphorylated to ATP using a high-energy
intermediate is referred to as a substrate-level
phosphorylation.
• In contrast to oxidative phosphorylation in
mitochondria, substrate-level phosphorylations
are not dependent on oxygen, and are the only
means of ATP generation in an anaerobic tissue.
• Pyruvate kinase—the last enzyme in aerobic
glycolysis, it catalyzes a substrate-level
phosphorylation of ADP using the high-energy
substrate phosphoenolpyruvate (PEP).
• Global regulation is mediated by insulin and
glucagon.
• Insulin stimulates and glucagon inhibits PFK-1
in hepatocytes by an indirect mechanism
involving PFK-2 and fructose 2,6-
bisphosphate.
• PFK-1 is inhibited by ATP and citrate, and
activated by AMP.
• Insulin activates PFK-2, which converts a tiny
amount of fructose 6-phosphate to fructose
2,6-bisphosphate (F2,6-BP). F2,6-BP activates
PFK-1.
• Glucagon inhibits PFK-2 (via cAMP-
dependent protein kinase A), lowering F2,6 BP
and thereby inhibiting PFK-1.
• Pyruvate kinase is activated by fructose 1,6-
bisphosphate from the PFK-1 reaction (feed-
forward activation).
• Hemolytic anemia results from pyruvate
kinase deficiency.
• Lactate dehydrogenase—is used only in anaerobic
glycolysis. It reoxidizes NADH to NAD+ by reducing
pyruvate to lactate, replenishing the oxidized
coenzyme for glyceraldehyde 3-phosphate
dehydrogenase.
• Without mitochondria and oxygen, glycolysis would
stop when all the available NAD+ had been reduced to
NADH.
• When oxygenation is poor in aerobic tissues (skeletal
muscle during strenuous exercise, myocardial
infarction), most cellular ATP is generated by
anaerobic glycolysis, and lactate production increases.
 Importance intermediate of Glycolysis

• Dihydroxyacetone phosphate (DHAP) is used

in liver and adipose tissue for triglyceride
synthesis.
• 1,3-Bisphosphoglycerate and
phosphoenolpyruvate (PEP) are high-energy
intermediates used to generate ATP by
substrate-level phosphorylation.
 .Regulation of Glycolysis
Glycolysis Is Regulated at Three Steps
Involving Nonequilibrium Reactions.
.Hormonal regulation
.Glycolysis in RBCs
.Fate of Pyruvate
.Pyruvate dehydrogenase complex
.Regulation of PDC
 .Clinical importance
1.Absence of aldolaseA(found in RBCs and
muscle) causes hemolytic anaemia.
2.Triose phosphate isomerase deficiency leads to
neonatal heamolytic anaemia.
3.Enolase is inhibited by fluoride. To prevent
ongoing glycolysis in a patient blood samples
collected for sensitive glucose tolerance
tests,blood collected in tubes containing
fluoride.
Mutations in glucokinase gene leads to Maturity.4 •
onset diabetes of the young(MODY) type2. Patients
.present nonprogressive hyperglycemia
Phosphofructokinase deficiency leads to.5 •
accumulation of glycogen in muscles.This causes
.haemolytic anaemia and muscle cramps
Deficiency of pyruvate kinase causes decreased.6 •
production of ATP from glycolysis.RBCs have
insufficient ATP for their membrane pumps and a
.haemolytic anaemia
6.Arsenite and mercuric ions react with the SH groups of lipoic
acid and inhibit pyruvate dehydrogenase, as well as dietary
deficiency of thiamin, allowing pyruvate to accumulate.
Nutritionally deprived alcoholics are thiamin-deficient and may
develop potentially fatal pyruvic and lactic acidosis.
Patients with inherited pyruvate dehydrogenase deficiency,
which can be due to defects in one or more of the components
of the enzyme complex, also present with lactic acidosis,
particularly after a glucose load. Because of its dependence on
glucose as a fuel, brain is a prominent tissue where these
metabolic defects manifest themselves in neurologic
disturbances.
The Tricarboxylic Acid Cycle,Electron Transport chain and
.shuttles
THE CITRIC ACID CYCLE PLAYS A
PIVOTAL ROLE IN METABOLISM
TWELVE ATP ARE FORMED PER TURN
OF THE CITRIC ACID CYCLE
.Regulation of citric acid cycle
.ETC
.Inhibitors of Electron Transports Chain(ETC)
 The Respiratory Chain Is a Proton Pump

• The Chemiosmotic Theory

• The chemiosmotic hypothesis (also known as the
Mitchell hypothesis) explains how the free energy
generated by the transport of electrons is used to
produce ATP from
• ADP +Pi by the electron transport chain
– 1. Proton pump: Electron transport is coupled to the
phosphorylationof ADP by the transport of
protons(H+) across the inner mitochondrial membrane
fromthe matrix to the space.
.ATP synthase
• Estimates suggest that for each NADH
oxidized, complex I translocates four protons and
complexes
• III and IV translocate 6 between them.
As four protons are taken into the mitochondrion for each
ATP exported, the P:O ratio would not necessarily be a
complete integer, ie, 3, but possibly 2.5.
However, for simplicity,
a value of 3 for the oxidation of NADH + H+
and 2 for the oxidation of FADH2 .
Inhibitors
• Oligomycin
• Uncoupling drugs-2,4 Dintrophenol.
.Glycerophosphate shuttle
.Malate shuttle
.The Creatine Phosphate Shuttle
 .Clinical importance
1.Fatal infantile mitochondrial myopathy and renal
dysfunction involves severe diminution or absence of
most oxidoreductases of the respiratory
chain(complexesI,III,IV and V).Patients present with early
progressive liver failure and neurologic
abnormalities,hypoglycemia and increased lactate in body
fluids.
2. MELAS (mitochondrial encephalopathy,
lactic acidosis, and stroke) is an inherited condition due to
NADH:ubiquinone oxidoreductase (complex I)
orcytochrome oxidase deficiency.
3.Leber,s hereditary optic neuropathy(LHON) is due to point
mutations in the gene for cytochrome reductase. Patients are
typically males in their 20s and 30s who develop loss of central
vision.
4.Kearns-Sayre syndrome have mutations in complexII and are
short stature, complete external ophthalmoplegia, pigmentary
retinopathy, ataxia and cardiac conduction defects.
5.Leigh disease is due to mutations in cytochrome oxidase. It is
fatal by age of 2yrs and present with lactic acidemia,
developmental delay, seizure, extraocular palsies and
hypotonia.