GLYCOLYSIS:

EMBDEN-MEYERHOF PATHWAY
The pathway for lactate fermentation in muscle is the same pathway as alcohol fermentation

ROLE OF GLUCOSE
Glucose is very soluble source of quick and ready energy. It is a relatively stable and easily transported. In mammals, the brain uses only glucose under non-starvation conditions.
Under starvation conditions ketone bodies become a source of energy for the brain

Glucose is the only source of energy in red blood cells.

GLYCOLYSIS
 The pathway for lactate fermentation in muscle is the same pathway as alcohol fermentation. It s showing an underlying unity in biology, i.e., oxidation of glucose into 2 molecules of pyruvic acid.  Expressed by: Gustav Embden Otto Meyerhof

Otto Meyerhof Nobel Prize 1922

GLYCOLYSIS
 The sequence of reactions from glucose to pyruvic acid is common to carbohydrate metabolism under both aerobic and anaerobic conditions.  In humans, under anaerobic conditions, the end product of glycolysis is two molecules of lactic acid.  Under aerobic conditions, the end product is two molecules of pyruvic acid.  Two molecules of ATP are produced; a limited amount.

GLYCOLYSIS
 Glycolysis takes place in the cytosol.  Under aerobic conditions the end-product is pyruvic acid.
Pyruvate is completely oxidized to CO2 in the TCA cycle and large amounts of ATP are subsequently produced.

 Under anaerobic conditions the end product is lactic acid; often referred to as lactic acid fermentation.  Some organisms (yeast) under anaerobic conditions convert pyruvate to ethanol; alcoholic fermentation.

The Fate of Glucose

Yeast

The fate of glucose is varies with physiological conditions, tissues, and organisms.

Exercising muscle

GLYCOLYSIS
Glycolysis devided into two stages: Stage 1 - Investment of ATP. Glucose is phosphorylated. The negative charge concentrates glucose in the cell and glucose becomes less stable. The 6 carbon sugar is split to two 3-carbon fragments. Stage 2 Energy yielding phase (harvesting of ATP or payoff). The oxidation of the 3-carbon fragments yields ATP

GLYCOLYSIS (Stage 1)

GLYCOLYSIS (Stage 2)

GLYCOLYSIS
First Stage of Glycolysis
Reaction 1: Phosphoryl transfer reaction 1. Kinases enzyme transfer phosphate group from ATP to an acceptor. Hexokinase has a more general specificity in that it can transfer phosphate to other sugars such as mannose. While glucokinase transfer phosphate group to glucose only.

GLYCOLYSIS
First Stage of Glycolysis
Reaction 2: Connversion of glucose 6-phosphate (an aldose) to fructose 6-phosphate (a ketose). This reaction is catalized by phosphoglucose isomerase

GLYCOLYSIS

First Stage of Glycolysis

Reaction 3: Phosphoryl transfer reaction 2 that convert fructose 6-phosphate to fructose 1,6-bisphosphate. In this step, the 2nd of ATP is invested. The reaction is catalized by phosphofructokinase-1 (PFK ).

PFK is an important allosteric enzyme regulating the rate of glucose catabolism and plays a role in integrating metabolism. Prefix bis means two phosphate groups on two different carbon atoms. What s the different with prefix di ?

GLYCOLYSIS

First Stage of Glycolysis

Reaction 4: Split of 6 carbon sugar of fructose 1,6bisphosphate to two 3-carbon sugar; glyceraldehyde 3phosphate and dihydroxyacetone phosphate. The reaction is reverse of aldol condensation that catalyzed by aldolase enzyme.

GLYCOLYSIS

First Stage of Glycolysis

Reaction 5: Isomerization of dihydroxyacetone phosphate to glyceraldehyde 3-phosphate . This reaction is catalized by triose phoshate isomerase enzyme.

(Go = 7,5 kJ/mol

All the DHAP is converted to glyceraldehyde 3-phosphate. This reaction is reversible, but glyceraldehyde 3-phosphate is a substrate for the next reactions of glycolysis. Thus, both 3carbon fragments are subsequently oxidized.

GLYCOLYSIS Second Stage of Glycolysis

Reaction 6: Oxidation of glyceraldehyde 3-phosphate to 1,3bisphosphoglycerat. In the reaction that catalyzed by glyceraldehyde 3-phosphate dehydrogenation enzyme, an aldehyde is oxidized to carboxylic acid and inorganic phosphate is transferred to form acyl-phosphate. NAD+ is reduced to NADH. 1,3-BPG is a mixed anhydride that has a high phosphoryl-transfer potential.

GLYCOLYSIS Second Stage of Glycolysis

Reaction 7: Transfer phosphate group from 1,3bisphosphoglycerate to ADP yielding an ATP. The reaction that catalyzed by phosphoglycerate kinase is a first substratelevel phosphorylation in glycolysis. In this reaction 1,3bisphosphoglycerate changes to 3-phosphoglycerate. At this point 2ATPs are produced.

GLYCOLYSIS Second Stage of Glycolysis

Reaction 8: Isomerization of 3-phosphoglycerate to 2phosphoglycerate. This is a phosphate shift reaction that catalyzed by phosphoglycerate mutase.

GLYCOLYSIS Second Stage of Glycolysis

Reaction 9: Dehydration of 2-phosphoglycerate to be phosphoenolpyruvate (PEP). This reaction is catalyzed by enolase.

GLYCOLYSIS Second Stage of Glycolysis

Reaction 10: Transfer phosphate group from phosphoenolpyruvate to ADP yielding an ATP. The reaction that catalyzed by pyruvate kinase is a second substrate-level phosphorylation in glycolysis. Phosphoenolpyruvate that unstable converted to pyruvate. At this point 2ATPs are produced.

GLYCOLYSIS Net Reaction of Glycolysis

Glucose + 2 Pi + 2 ADP + 2 NAD+ 2 pyruvate + 2 ATP + 2 NADH +2 H+

REGULATION OF THE GLYCOLYTIC PATHWAY

Enzyme reactions that have a significant negative G are often control sites. Regulatory enzymes in glycolysis are:
 Hexokinase  Phosphofructokinase (PFK)  Pyruvate Kinase
PFK is the most important regulatory enzyme.

REGULATION OF THE GLYCOLYTIC PATHWAY

Hexokinase Regulates Glycolysis
 Hexokinase is inhibited by its product, glucose 6phosphate.  High concentrations of glucose 6-phosphate indicates that the cell no longer needs glucose for energy, instead glucose for storage as glycogen, or for other precursors.  Remember that the liver is responsible for regulating blood glucose levels.

REGULATION OF THE GLYCOLYTIC PATHWAY

Hexokinase and Glucokinase
 The liver contains an isoform of hexokinase called glucokinase Glucokinase is not inhibited by glucose 6phosphate. Glucokinase has a lower affinity for glucose than hexokinase.
This assures that brain and muscle have first choice for the glucose.

When glucose is abundant in the liver, glucokinase phosphorylates glucose to glucose 6phosphate specifically for glycogen synthesis.

REGULATION OF THE GLYCOLYTIC PATHWAY Phosphofructokinase is an Allosteric Enzyme

High levels of ATP inhibit PFK. Increased levels of AMP reverses the action of ATP. Citrate also inhibits PFK. High levels of citrate indicates that the cell is rich in biosynthetic precursors.

Fructose 2,6-bisphosphate is a Positive Effector for PFK

F-2,6-BP increase affinity of PFK to fructose 6-phosphate. Thus, stimulating glycolysis.

Note: An allosteric protein is one in which the binding of a ligand to one site affects the binding properties of another site on the same protein.

REGULATION OF THE GLYCOLYTIC PATHWAY

Pyruvate Kinase Has Regulatory Role in Glycolysis  Pyruvate kinase has an L (liver) and M (muscle and brain) form.  Both forms are inhibited by its product, ATP.  Fructose 1,6-bisphosphate activates both forms of the enzyme to keep pace with the influx (supaya selaras dengan arus) on intermediates.  Alanine can be reversibly transaminated to pyruvate. Alanine also inhibits pyruvate kinase thus indicating that building blocks are abundant.

REGULATION OF THE GLYCOLYTIC PATHWAY Phosphorylation of Pyruvate Kinase

When blood-glucose levels are low the glucagon phosphorylates pyruvate kinase making it less active.

Pyruvate kinase: only L-form is regulated by covalent modification.

HOW ARE OTHER SUGARS FEED INTO THE GLYCOLYTIC PATHWAY ?

FRUCTOSE FEED INTO THE GLYCOLYTIC PATHWAY

In the liver, when fructose enters glycolysis the PFK reaction is bypassed. Fructose metabolism in the liver

GALACTOSE FEED INTO THE GLYCOLYTIC PATHWAY

Galactose is converted to glucose-6P via a four step reaction involving UDPglucose Please, look for four step reaction converting galactose to glucose-6 phosphate !

Conversion of galactose to glucose 1phosphate. The conversion proceeds through a sugarnucleotide derivative, UDP-galactose, which is formed when galactose 1-phosphate displaces glucose 1-phosphate from UDP-glucose. UDP-galactose is then converted by UDPglucose 4-epimerase to UDP-glucose.

GLUCONEOGENESIS
Gluconeogenesis is the synthesis of glucose from non-carbohydrate precursors. Glucose stores are depleted during periods of starvation or fasting beyond a day. Since the brain relies on glucose (120g/d) as a source of energy, glucose must be synthesized from molecules other than carbohydrates. PYRUVATE GLUCOSE

GLUCONEOGENESIS
Any molecule that can be converted to pyruvate is considered glucogenic. Lactate and alanine are glucogenic. Glycerol is also glucogenic.

GLUCONEOGENESIS
The enzymes in red belong to the gluconeogenic pathway. These reactions overcome the high negative free energy of the irreversible reactions of glycolysis. The enzymes in blue are held in common between the two pathways.

GLUCONEOGENESIS
 The irreversible glycolytic enzymes are: hexokinase ( G =-8 kcal mol-1), phosphofructokinase ( G = -5.3 kcal mol-1 ), and pyruvate kinase ( G = -4.0 kcal mol-1).  The enzymes of gluconeogenesis are:
   

pyruvate carboxylase (ATP) phosphoenolpyruvate carboxykinase (GTP) fructose 1,6-bisphosphatase glucose 6-phosphatase

GLUCONEOGENESIS
Pyruvate Carboxylase
 Pyruvate + CO2 + ATP + H2O p oxaloacetate + ADP + Pi + 2 H+  Pyruvate Carboxylase fixes CO2. Enzymes which fix CO2 require the cofactor BIOTIN. Biotin is a vitamin and is always involved in CO2 fixation.  This reaction takes place in the mitochondrial matrix.

GLUCONEOGENESIS
Biotin

GLUCONEOGENESIS
Phosphoenolpyruvate Carboxykinase
 Oxaloacetate + GTP p phosphoenolpyruvate + GDP + CO2  This reaction takes place in the cytosol  PEP is now synthesized from pyruvate and the sum of the two reaction is: Pyruvate + ATP + GTP + H2O p PEP + ADP + GDP + Pi + H+

GLUCONEOGENESIS
Pyruvate --- phosphoenolpyruvate
Pyruvate is carboxylated in the mitochondria by pyruvate carboxylase. Oxaloacetate can t pass out of the mitochondria. It is converted to malate by malate dehydrogenase and pass out of the mitochondria. In the cytosol malate hydrogenated again back to oxaloacetate . Finaly, oxaloacetate decarboxylated and phosphorylated to be phosphoenolpyruvate by phosphoenolpyruvate carboxykinase .

Malate DH

Malate DH

GLUCONEOGENESIS

Fructose 1,6-bisphosphatase
 Fructose 1,6-bisphosphate + H2O p fructose 6-phosphate + Pi  Fructose 1,6-bisphosphatase is an allosteric enzyme and regulates gluconeogenesis.  Fructose 6-phosphate is easily converted to glucose 6-phosphate.

GLUCONEOGENESIS
Glucose 6-phosphatase
 Glucose 6-phosphate + H2O glucose + Pi.  Liver can send glucose to blood to maintain homeostasis.  Glucose 6-phosphate is also a precursor to glycogen. Note:
 Homeostasis: the maintenance of a stable equilibrium, especially through physiological processes.

GLUCONEOGENESIS
End of Gluconeogenesis
 Gluconeogenesis ends at glucose 6-phosphate.  Glucose 6-P is valuable; a precursor for glycogen synthesis.  Glucose 6-phosphatase is present only in tissues responsible for maintaining blood glucose levels, liver and kidney.  In liver, glucose 6-phosphatase is highly regulated.

NET REACTION OF GLUCONEOGENESIS

2pyruvate + 4ATP + 2GTP + 2NADH + 6H2O p glucose + 4ADP + 2GDP +6Pi +2NAD+ + 2H+ Note: it takes 6 nucleoside triphosphate molecules to synthesize glucose. Only 2 nucleoside triphosphate molecules are generated from glycolysis. So it takes four extra high phosphoryltransfer potential molecules to drive the unfavorable gluconeogenesis pathway.

GLYCOLYSIS VS GLUCONEOGENESIS
 Gluconeogenesis is the synthesis of glucose from non-carbohydrate precursors such as pyruvic and lactic acid.  Both pathways are not just the reverse of each other although some enzymes are held in common.  Specific reactions to gluconeogenesis bypass the highly exergonic reactions ( G <<0) of glycolysis.  The two pathways have distinctive regulatory mechanisms.

REGULATION OF GLYCOLYSIS AND GLUCONEOGENESIS

Glycolysis and gluconeogenesis are reciprocally regulated. A high [AMP] indicates that the energy charge is low and signals the need for ATP. High [ATP] and [citrate] indicate the energy charge is high and intermediates are abundant.
Insulin stimulates Glucagon stimulates

SYNTHETIC AND DEGRADATIVE PATHWAYS

Synthetic and degradative pathways are distinct: If [ATP] is low, degradative pathways are stimulated. If [ATP] is high, degradative pathways are inhibited.
Degradation

Synthesis

SYNTHETIC AND DEGRADATIVE PATHWAYS

Regulation of the degradation and synthesis of glucose and glycogen depends on the energy state of the cell: High [NADH] is indirectly equivalent to high[ATP]. This means that the cell is high in energy . High [NAD+] or [ADP or AMP] means that the cell is low in energy . These molecules (and others) can act as allosteric effectors stimulating or inhibiting allosteric enzymes which are usually at the beginning or branch-points of a specific pathway.

SYNTHETIC AND DEGRADATIVE PATHWAYS

Synthetic and degradative pathways don t happen at the same time:  They can share some common steps but they are never simply the reverse of one another.  Synthetic pathways always use more ATP than a degradative pathway will produce.  If both synthetic and degradative pathways occurred at the same time, wasteful hydrolysis of ATP would result.  This is termed a futile cycle.

THE FUTILE CYCLE

Fruc. 6-P + ATP Fruc. 1,6-BisP NET: ATP Fruc. 1,6-BisP Fruc. 6-P + Pi

ADP + Pi

To minimize futile cycling, signals that activate a synthetic pathway, inactivate the corresponding degradative pathway.

SYNTHETIC AND DEGRADATIVE PATHWAYS

 For example, phosphorylation activates glycogenolysis whereas phosphorylation inactivates glycogenesis (glycogen synthesis).  Put differently: Phosphorylation activates glycogenolysis whereas dephosphorylation activates glycogenesis.  On the same theme, the action of insulin is opposite to that of glucagon.  Insulin decreases blood glucose levels whereas glucagon increases blood glucose levels.

REGULATION OF BLOOD GLOCOSE