Glycolysis (Compatibility Mode)

Carbohydrate Metabolism
Sources Initial Absorption and Digestion Oxidation of Glucose: Glycolysis & Citric Acid Cycle Synthesis of Glucose: Gluconeogenesis Glycogen Metabolism Pentose Phosphate Pathway Organ Integration of Carbohydrate Metabolism
9/28/2012
Dr.Santosh Kumar
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
Dietary Sources of Carbohydrates

(30) (1) -1,4 & -1,6 glycosidic linkages 1 Branched 1 4 2 1 4 3 glu 1 2 fru 6 1
(Most abundant biomolecules on earth)
Starch: the nutritional store of glucose in plants. A major source of dietary glucose.
Similar to glycogen in animal cells, but less extensive branching in starch.
2 Forms of Starch (nutritional reservoir in plants)

-1,4 glycosidic linkages glu
glu
Unbranched
Disaccharides
glu
glu
gal
glu
Monosaccharides
gal glu fru
9/28/2012 Dr.Santosh Kumar 2 Carbohydrate oxidation is the central energy yielding pathway in most cells.
Glycolysis (Embden-Meyerhof Pathway)

1) Represents the central pathway for the catabolism of glucose and other monosaccharides.
2) A nearly universal pathway in biological systems and occurs in virtually every tissue.
3) The liver is the principal organ in the maintenance of blood glucose levels. Thus the functional state of the liver will profoundly influence carbohydrate metabolism.
4) D-glucose is oxidized to pyruvate, which under anaerobic conditions may then be partially converted to lactate.
9/28/2012
Dr.Santosh Kumar
Under anaerobic conditions: glycolysis consists of 11 coupled reactions with the overall net reaction being: D-glucose + 2 Pi + 2 ADP G = -32.4 kcal/mol 2 lactate + 2 H+ + 2 ATP + 2 H2O
Under aerobic conditions: glycolysis consists of 10 coupled reactions with the overall net reaction being: D-glucose + 2 Pi + 2 ADP + 2 NAD+ 2 pyruvate + 2 H+ + 2 ATP + 2 NADH + 2 H2O
G = -20.4 kcal/mol
9/28/2012 Dr.Santosh Kumar 4
Glycolysis can be divided into 2 stages:
Stage 1 (The Preparatory Phase): Consists of the collection of sugars by the liver. Subsequent phosphorylation of these sugars at the expense of 2 ATPs. Conversion to glyceraldehyde 3-phosphate.
Stage 2 (The Payoff Phase): Consists of conversion of glyceraldehyde 3-phosphate to either pyruvate or lactate via a series of oxidationreduction steps. Concomitant conservation of energy via formation of 4 ATPs (5 or 6 enzymes are utilized, respectively).
Stage 1 -2 ATP
Stage 1 -2 ATP
Stage 2 +4 ATP
Stage 2 +4 ATP
2 NAD+
2 NAD+ 11 Lactate (2)
9/28/2012
Dr.Santosh Kumar
Glycolysis: Stage 1
Initial Strategy: Trap glucose in the cell and convert it to a compound that can be cleaved into phosphorylated 3-carbon units. 1st Reaction: Glucose enters cell and is phosphorylated to glucose 6-phosphate, a negatively charged molecule which is trapped inside the cell.
9/28/2012
Dr.Santosh Kumar
Important Additional Points

1) Reaction is irreversible.
2) Liver also contains a specialized form of hexokinase known as glucokinase.
3) Glucokinase: much higher Km for glucose (5 mM vs. 0.1 mM) is not inhibited by glucose 6-phosphate is absent in muscle and is deficient in patients with diabetes
4) At normal blood glucose concentrations hexokinase is fully saturated, glucokinase is not.
5) Glucokinase is present at high concentration in liver and is induced in response to D-glucose.

6) Glucokinase assures that at high concentrations, glucose is not wasted. Instead it is converted to glucose 6-phosphate for subsequent synthesis of glycogen.
7) Hexokinase can also phosphorylate fructose, mannose, and glucosamine, whereas glucokinase cannot. However, due to its high Km for fructose, and existence of a separate pathway for fructose, it is not a significant substrate for hexokinase.
8) First reaction does not commit glucose to glycolysis, since glucose-6-phosphate represents a branch point in carbohydrate metabolism. It also enters pentose phosphate pathway and glycogenesis.
2nd Reaction: the isomerization of glucose 6-phosphate to fructose 6-phosphate. Catalyzed by phosphoglucose isomerase.
6 membered pyranose ring

6 5 4 3 2 1
5 membered furanose ring
1 2
4
5 3
This reaction is readily reversible.
9/28/2012
Dr.Santosh Kumar
10
This reaction represents an example of a conversion of an aldose to a ketose.
Open chain representation of the sugars.

1 2 3 4 5 6 Aldehyde
1 2 3 4 5 6 Ketone
An aldehyde containing sugar.
A ketone containing sugar.
9/28/2012
Dr.Santosh Kumar
11
3rd Reaction: Fructose 6-phosphate is phosphorylated by ATP to form fructose 1,6-bisphosphate. This is the second of the two priming reactions in glycolysis. Catalyzed by phosphofructokinase (PFK; PFK1).
6 5 4 3
1 2
PFK is the major point of regulation in glycolysis. Rx is irreversible. PFK is regulated allosterically.
4th Reaction: Cleavage of fructose 1,6-bisphosphate to glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. Represents cleavage of a hexose into two trioses.
1 2 3 4 5 6
1 2 3 4 5 6
Note: Reaction is readily reversible. It is pulled to the right via removal of glyceraldehyde 3-phosphate via subsequent steps. Only glyceraldehyde 3-phosphate is on the direct pathway of glycolysis.
5th Reaction: Isomerization of 3-carbon phosphorylated sugars. catalyzed by triose phosphate isomerase.
1 2 3
4 5 6
9/28/2012
Dr.Santosh Kumar
14
Additional Points: 1) At equilibrium 96% of triose phosphate is dihydroxyacetone phosphate. Rx. is pulled to the right via rapid removal of the product.
2) Net result: Dihydroxyacetone phosphate is funneled into the main glycolytic pathway; 2 molecules of glyceraldehyde 3-phosphate are formed from 1 molecule of fructose 1,6-bisphosphate.
3) Triose phosphate isomerase accelerates isomerization by a factor of 1010. Only limited by diffusion of the substrate into the active site. Considered a kinetically perfect enzyme.
9/28/2012
Dr.Santosh Kumar
15
With this reaction, carbons 1, 2, and 3 of the starting glucose become indistinguishable from carbons 6, 5, and 4, respectively. Also, the numbering of carbon atoms in glyceraldehyde 3-phosphate is not the same as the numbering of carbons in glucose.
9/28/2012
Dr.Santosh Kumar
16
Glycolysis: Stage 2
Stage 1: 1 molecule of glucose 3-phosphate. 2 molecules of glyceraldehyde
Stage 2: 2 molecules of glyceraldehyde 3-phosphate 2 molecules of pyruvate AND 4 ADP 4ATP.
9/28/2012
Dr.Santosh Kumar
17
Stage 1 -2 ATP
Stage 1 -2 ATP
Stage 2 +4 ATP
2 NAD+ 2 NAD+
2 NAD+ 11 Lactate (2)
9/28/2012
Dr.Santosh Kumar
18
6th Reaction: Oxidation of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate.

1 2 3
Aldehyde group
NAD+dependent
Note: The mixed anhydride has a very high free energy of hydrolysis.
1 2 3
Mixed anhydride This is a mixed anhydride of phosphoric acid and a carboxylic acid.
9/28/2012
The first of the two energy-conserving reactions of glycolysis that will ultimately Dr.Santosh yield ATP. Kumar
19
7th Reaction: First ATP-generating step. ATP is formed as the phosporyl on the carboxyl group of 1,3-bisphosphoglycerate is transferred to ADP. Substrate Level Phosphorylation
1 2 3
Note: The consequences of this reaction in combination with the 6th reaction are: 1) An aldehyde is oxidized to a carboxylic acid group. 2) NAD+ is concomitantly reduced to NADH.
1 2 3
3) ATP is formed from Pi and ADP.
9/28/2012
Dr.Santosh Kumar
20
8th Reaction: The phosphoryl group is shifted from the C-3 to the C-2 position of glycerate. Catalyzed by phosphoglycerate mutase. Note: A mutase transfers a functional group from one position to another on the same molecule.
1 2 3
1 2 3
9/28/2012
Dr.Santosh Kumar
21
9th Reaction: A dehydration reaction is which water is reversibly removed from 2-phosphoglycerate to from phosphoenolpyruvate. Catalyzed by enolase. Large difference in the standard free energy of hydrolysis of the phosphate group in the reactant versus the product.
1 2 3
1 2 3
9/28/2012
Dr.Santosh Kumar
22
10th Reaction: Transfer of a phosphoryl group from PEP to ADP catalyzed by pyruvate kinase. Irreversible; An important site of regulation in the liver. The second substrate level phosphorylation.
9/28/2012
Dr.Santosh Kumar
23
11th Reaction: Reduction of pyruvate to lactate via the enzyme lactate dehydrogenase. Conversion of occurs under partially anerobic conditions, when oxygen is limited (e.g., muscle during intense activity) OR in certain tissues even when sufficient oxygen is present (retina, brain, RBCs).
NADH required for this reaction is supplied by the 6th reaction (the dehydrogenation of glyceraldehyde 3-phosphate).
Importantly, under anaerobic conditions, the regeneration of NAD+ by this step is essential for the continued functioning of glycolysis.
9/28/2012
Dr.Santosh Kumar
24
Stage 1 -2 ATP
Stage 2 +4 ATP
2 NAD+ 11
9/28/2012
Dr.Santosh Kumar
Lactate (2)
25
There are 5 isozymic forms of lactate dehydrogenase. Differ in their affinity for substrate and sensitivity to allosteric inhibition. Isozymes: multiple forms of a given enzyme that catalyze the same reaction but differ in kinetic or regulatory properties. Each LDH isozyme contains 4 copes of two different polypeptides. H form (heart) and M form (muscle). Designated H4, H3M1, H2M2, etc. H4: higher affinity for substrate; allosterically regulated. Designed to oxidize lactate to pyruvate which can be used by the heart as an aerobic fuel source. M4: optimized to convert pyruvate to lactate in muscle; allows glycolysis to continue under anaerobic conditions.
Net reaction in transformation of glucose into pyruvate: D-glucose + 2 Pi + 2 ADP + 2 NAD+ 2 pyruvate + 2 ATP + 2 NADH + 2 H+ + 2 H2O
2 ATPs are generated during conversion of glucose to 2 pyruvate.
3 Reactions are irreversible under physiological conditions: hexokinase phosphofructokinase pyruvate kinase
9/28/2012
Dr.Santosh Kumar
27
Entry of Other Monosaccharides into Glycolysis
9/28/2012
Dr.Santosh Kumar
28
D-Fructose: present in fruits; also can be generated by hydrolysis of sucrose (yield fructose + glucose). Note: In the liver hexokinase has a 20x higher affinity for glucose compared to fructose. Since there is a lot of glucose present in this organ, fructose is not principally metabolized by hexokinase, but rather by the following pathway:
9/28/2012
Dr.Santosh Kumar
29
(2) (1)
(1) Fructose intolerance results from a deficiency in fructose 1-phosphate aldolase. Leads to an accumulation in fructose 1-phosphate and a depletion of ATP and Pi. Pi depletion makes it impossible to generate more ATP lowering levels even further. Causes cell damage. (2) Fructosuria results from a deficiency in fructokinase. Fructose appears in blood and urine. Relatively benign metabolic abnormality.
9/28/2012
Dr.Santosh Kumar
31
D-mannose: arises from digestion of polysaccharides and glycoproteins found in food.
Hexokinase
Mannose + ATP
Mannose 6-phosphate
Phosphomannose isomerase
Mannose 6-phosphate
Fructose 6-phosphate
9/28/2012
Dr.Santosh Kumar
32
D-Galactose: Derived via hydrolysis of the disaccharide lactose. It is converted to glucose 1-phosphate as follows: Step 1: galactose is phosphorylated.
Step 2: Galactose 1-phosphate acquires a uridyl group from UDP-glucose with a release of glucose 1-phosphate.
UDP
Step 3: The galactose moiety of UDP-galactose is epimerized to glucose; the configuration of the hydroxyl group at C-4 is inverted. Net effect is conversion of galactose 1phosphate to glucose 1-phosphate.
Glucose 1-phosphate is then isomerized to glucose 6-phosphate:

phosphoglucomutase
Glucose 1-phosphate
Glucose 6-phosphate
The disease galactosemia results from the absence of the enzyme galactose 1-phosphate uridyl transferase. Galactose metabolism is blocked at the galactose 1-phosphate step. Damage occurs due to the accumulation of toxic substances rather than due to the absence of an essential compound.
9/28/2012
Dr.Santosh Kumar
34
One of the offending compounds is galactitol which is produced by the reduction of galactose. Galactosemia is a severe disease. Symptoms occur when milk is consumed; liver becomes enlarged; jaundice is common. Blood galactose is elevated and its found in the urine is well. Absence of the transferase enzyme from RBCs is diagnostic. Treatment involves exclusion of galactose from the diet.
Conversion of Glucose to Fructose via Sorbitol

Aldose reductase reduces glucose to sorbitol, which is quite polar and thus does not passively diffuse across membrane. Also, its not a substrate for the glucose transporter. Therefore its trapped inside cells.
Liver, ovaries, sperm, and seminal vesicles contain the enzyme sorbitol dehydrogenase. Oxidizes sorbitol to fructose. Fructose then enters glycolysis or gluconeogenesis.
9/28/2012
Dr.Santosh Kumar
36
When glucose is elevated (e.g., diabetes) and if there is sufficient NADPH, aldose reductase produces excess sorbitol. Retina, lens, kidney, and nerve cells do not contain sorbitol dehydrogenase and therefore sorbitol accumulates. Causes a strong osmotic effect and cell swelling due to water retention. Symptomalogy occurs (cataract formation, peripheral neuropathy, and vascular problems).
No sorbitol dehydrogenase.
9/28/2012
Dr.Santosh Kumar
37
Dietary Disaccharides are Hydrolyzed to Monosaccharides

Disaccharides cannot directly enter cells without first being hydrolyzed to monosaccharides (extracellularly). Hydrolysis reactions are catalyzed by enzymes attached to outer surface of epithelial cells lining the small intestine.
The resulting monosaccharides enter cells lining the intestine via specific transport proteins. Then pass from cells into the blood, distributed to the liver, enter the glycolytic pathway.
Regulation of Glycolysis
Enzymes catalyzing irreversible reactions are often potential control sites. In glycolysis regulation occurs at hexokinase, phosphofructokinase, and pyruvate kinase. I. Phosphofructokinase: the most important control point in glycolysis. It is the first irreversible reaction that is unique to the pathway (i.e., the committed step). Inhibited by ATP AMP reverses the inhibition by ATP. Thus PFK activity increases when the ATP/AMP ratio is lowered (i.e., as the energy charge of the cell decreases). Inhibited by a decrease in pH. Inhibited by citrate.
9/28/2012
Activated by Fructose 2,6-bisphosphate.*** Dr.Santosh Kumar
39
Fructose 2,6-BP is formed by phosphorylation of fructose 6-phosphate via the enzyme phosphofructokinase 2. Fructose 2,6-BP can be hydrolyzed by the enzyme fructose bisphosphatase 2. Fructose 6-phosphate both accelerates the synthesis of fructose 2,6-BP and inhibits its hydrolysis. Both mechanisms lead to increased fructose 2,6-BP.
Phosphofructokinase 2
Causes phosphorylation of the enzyme which then activates the phosphatase function.
Fructose bisphosphatase 2
9/28/2012
Dr.Santosh Kumar
40
The activities of PFK2 and FBPase2 reside on the same polypeptide chain. Both activities are reciprocally regulated by phosphorylation of a single serine residue.
Thus low blood glucose, which then
blood glucagon,
cAMP-dependent PFK2 and FBPase 2,
phosphorylation of this bifunctional enzyme, F 2,6-BP, and
the activity of PFK1.
9/28/2012
Dr.Santosh Kumar
41
Phosphofructokinase 2
Causes phosphorylation of the enzyme which then activates the phosphatase function and inhibits the kinase.
Fructose bisphosphatase 2
Net result is to decrease fructose 2,6-bisphosphate level and therefore decrease PFK1 activity.
9/28/2012
Dr.Santosh Kumar
42
II. Hexokinase: inhibited by its product glucose 6-phosphate. Thus PFK inhibition leads to hexokinase inhibition via the buildup of metabolites.
III. Pyruvate Kinase: catalyzes the third irreversible step in glycolysis. Controls the outflow of the pathway. 2 forms: L form (liver): subject to extensive allosteric regulation. Fructose 1,6-bisphosphate activates. ATP and alanine inhibit.
M form (muscle and brain): not allosterically regulated.

The L form of pyruvate kinase is inhibited by hormone-mediated cAMP-dependent phosphorylation as depicted below:
Low glucose inhibits dephosphorylation of pyruvate kinase thereby maintaining the inactive form of the enzyme.
Low glucose stimulates phosphorylation of pyruvate kinase which inactivates the enzyme.
Low blood glucose, glucagon, cAMP-dependent phosphorylation, of pyruvate kinase, INACTIVATES. Thus low blood glucose, PFK1 and pyruvate kinase.
Bottom Line: liver does not consume glucose when it is more urgently needed by9/28/2012 brain and muscle. Dr.Santosh Kumar 44
Alcoholic Fermentation
The sequence of reactions from glucose to pyruvate is similar in all organisms. However, in yeast and several other microorganisms ethanol is formed from pyruvate via the following 2 reactions: Note: the CO2 produced via pyruvate decarboxylation in Brewers yeast is responsible for the carbonation of champagne. In baking the CO2 released when yeast is mixed with a fermentable sugar causes the dough to rise.
Present in humans. partly responsible for the oxidation of ethanol.
Absent in Humans.
9/28/2012
Dr.Santosh Kumar
45

Glycolysis (Compatibility Mode)

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Glycolysis (Compatibility Mode)

Uploaded by

Copyright:

Available Formats

Carbohydrate Metabolism

Dietary Sources of Carbohydrates

(Most abundant biomolecules on earth)

Similar to glycogen in animal cells, but less extensive branching in starch.

2 Forms of Starch (nutritional reservoir in plants)

Glycolysis (Embden-Meyerhof Pathway)

Glycolysis can be divided into 2 stages:

2 NAD+ 11 Lactate (2)

Important Additional Points

2) Liver also contains a specialized form of hexokinase known as glucokinase.

4) At normal blood glucose concentrations hexokinase is fully saturated, glucokinase is not.

5) Glucokinase is present at high concentration in liver and is induced in response to D-glucose.

6 membered pyranose ring

5 membered furanose ring

This reaction is readily reversible.

This reaction represents an example of a conversion of an aldose to a ketose.

Open chain representation of the sugars.

An aldehyde containing sugar.

A ketone containing sugar.

Stage 2: 2 molecules of glyceraldehyde 3-phosphate 2 molecules of pyruvate AND 4 ADP 4ATP.

2 NAD+ 11 Lactate (2)

6th Reaction: Oxidation of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate.

3) ATP is formed from Pi and ADP.

2 ATPs are generated during conversion of glucose to 2 pyruvate.

Entry of Other Monosaccharides into Glycolysis

D-mannose: arises from digestion of polysaccharides and glycoproteins found in food.

Glucose 1-phosphate is then isomerized to glucose 6-phosphate:

Conversion of Glucose to Fructose via Sorbitol

Dietary Disaccharides are Hydrolyzed to Monosaccharides

Activated by Fructose 2,6-bisphosphate.*** Dr.Santosh Kumar

Thus low blood glucose, which then

cAMP-dependent PFK2 and FBPase 2,

phosphorylation of this bifunctional enzyme, F 2,6-BP, and

the activity of PFK1.

M form (muscle and brain): not allosterically regulated.

You might also like