Pathophysiology 101-823

Unit 3 Metabolism & Metabolic Disease

Carbohydrate Metabolism
PAUL ANDERSON 2008

Learning Objectives
1. Describe the metabolic functions of carbohydrates. 2. Describe the following pathways of Cell Respiration: for each state

the type of reactions & their functions, compare aerobic vs anaerobic respiration, cellular locations. - Glycolysis - Krebs (TCA) Cycle - Oxidative Phosphorylation 3. Describe Gluconeogenesis and its hormonal controls. 4. Describe the following processes in liver and muscle. - Glycogenesis - Glycogenolysis 5. Define Glycogen Storage Diseases and predict the biochemical consequences of enzyme deficiencies in the pathways listed. 6. Define Mitochondrial Disease and explain the symptoms.
References: Martini, Essentials of Anatomy & Physiology, Ch. 17 Porth, Essentials of Pathophysiology, Ch. 4, p.67 (mitochondrial disease), Ch. 32 Glucose Metabolism Or Porth, Pathophysiology (hard cover), Ch. 7, pp. 140-141 (mitochondrial disease), Ch. 43 Glucose Metabolism

Functions of Carbohydrates -1
Carbohydrates provide "IMMEDIATE" ENERGY for cells. Blood sugar (dextrose or glucose) is the "fuel molecule" used by all cells (especially brain cells) for ATP synthesis. Energy is released for biological work when glucose is oxidised in the process of aerobic cellular respiration (ATP temporarily stores the energy).
glucose biological work ATP

ADP + Pi

Functions of Carbohydrates -2
Carbohydrates provide STORED ENERGY as glycogen (mainly in liver, muscles) Hepatic glycogen acts as a reservoir for the blood sugar. Blood glucose levels are controlled by hormones which affect Gycogen storage. Hypoglycemic hormone

insulin

glucose +

glycogen

epinephrine glucagon

Hyperglycemic hormones

Pathways of Carbohydrate Metabolism

Cell Respiration - Glycolysis - Krebs (TCA) Cycle - Oxidative Phosphorylation Gluconeogenesis Glycogenesis Glycogenolysis

CELL RESPIRATION
CELL RESPIRATION is the process of generating ATP through the energy released from OXIDATION of organic compounds. CELL RESPIRATION in AEROBIC CELLS involves three metabolic pathways - GLYCOLYSIS - the KREBS CYCLE (TCA or Citric Acid Cycle) - OXIDATIVE PHOSPHORYLATION. The KREBS CYCLE and OXIDATIVE PHOSPHORYLATION require OXYGEN and occur in the MITOCHONDRIA. GLYCOLYSIS occurs in the CYTOSOL and can occur in the presence or absence of oxygen. Cell Respiration begins with glycolysis: the usual starting point is glucose.

CELL RESPIRATION Summary Equation

In

Cell Respiration

Carbon in Glucose loses electrons (in H atoms) and becomes oxidized to CO2 O2 gains electrons (in H atoms) and becomes reduced to H2O
Along the way, the electrons lose potential energy, when they fall from organic compounds to oxygen during cellular respiration and energy is released to form ATP.

CELL RESPIRATION
CELL RESPIRATION consists of two COUPLED REACTIONS, the EXERGONIC OXIDATION of GLUCOSE and the ENDERGONIC SYNTHESIS of ATP.

C6H12O6 + 6O2 6CO2 + 6H2O + ENERGY

38ATP + 38H2O <----- 38ADP + 38Pi + ENERGY

The maximum possible ATP yield AEROBICALLY is 38 ATP per GLUCOSE or 40% of the energy released from glucose oxidation, with the remaining 60% of energy escaping as heat.

CELL RESPIRATION
Oxidation of glucose does not occur in one reaction but in a series of smaller DEHYDROGENATION reactions, each of which releases a small amount of energy. DEHYDROGENATIONS remove ELECTRONS from a compound and so are also OXIDATIONS. OXIDATIONS always occur with REDUCTION of another compound (which accepts HYDROGEN or ELECTRONS). The two half reactions make up one REDOX REACTION. Whereas OXIDATION is EXERGONIC REDUCTION is ENDERGONIC and so the two half reactions are COUPLED.

Note: Electrons lose energy as they flow from XH2 to an oxidising agent, Y.

CELL RESPIRATION

When ELECTRONS are transferred to an OXIDISING AGENT they are moving to a lower energy state. The energy difference appears as released free energy which can be used to form ATP. CELL RESPIRATION consists of a series of these REDOX REACTIONS which release energy for ATP SYNTHESIS.

Electron Flow in Cell Respiration Releases Energy for ATP Synthesis

NADH NAD+
2e H+

ATP Controlled release of energy for synthesis of ATP

El H2 is transferred by a ec tro coenzyme (usually NAD) n ch tran to carriers on an electron ain sp or transport chain. t Redox reactions on the chain release energy for ATP synthesis. Oxygen,the final H2 acceptor and most powerful oxidising agent is reduced to H2O.

2e 2 H+
1 2

O2

H2O

Oxidative Phosphorylation & The Electron Transport Chain

OXIDATIVE PHOSPHORYLATION is the indirect way of generating ATP from ADP and Pi using the energy released from REDOX REACTIONS on the ELECTRON TRANSPORT (RESPIRATORY) CHAIN. The ELECTRON TRANSPORT CHAIN is a series of compounds on the INNER MITOCHONDRIAL MEMBRANE which transfers H2 or electrons from one compound to another in a series of REDOX REACTIONS.

The Electron Transport Chain - 1

The ELECTRON TRANSPORT CHAIN is a series of increasingly more powerful OXIDISING AGENTS beginning with REDUCED NAD (NADH) and ending with OXYGEN. As ELECTRONS pass along the chain they are moving to lower and lower energy levels. Each member of the chain is alternately REDUCED (when it receives ELECTRONS) and then OXIDISED (when it loses ELECTRONS). When reduced each compound acts as a REDUCING AGENT for the next member of the chain.

The Electron Transport Chain - 2

The first member of the ELECTRON TRANSPORT CHAIN is NADH (reduced Nicotinamide Adenine Dinucleotide) which contains the vitamin Nicotinic Acid (Niacin). NAD picks up hydrogens from compounds of GLYCOLYSIS and the KREBS CYCLE and transfers them to a Flavoprotein, the next member of the chain on the inner mitochondrial membrane. Flavoprotein is a conjugated protein consisting of an enzyme complex (NADH dehydrogenase) containing a non - protein FMN (Flavin Mononucleotide) which receives and passes on H2 NADH + H+ + FMN --------> NAD+ + FMNH2

The Electron Transport Chain - 3

A lipid (COENZYME Q or UBIQUINONE) takes ELECTRONS from FMN and transfers them to CYTOCHROMES (PROTEIN pigments). The last CYTOCHROME is CYTOCHROME OXIDASE which transfers ELECTRONS to OXYGEN reducing it to H2O. The energy drop for electrons passed from NADH to H2O is enough to generate a maximum of 3 ATP per NADH molecule. HYDROGENS are also delivered to the chain by FADH2. REDUCED FAD (Flavin Adenine Dinucleotide) enters the chain at a lower level than NADH so generates only 2 ATP per FADH2.

The Electron Transport Chain - 4

Glycolysis Overview
GLYCOLYSIS the first stage of CELL RESPIRATION Glucose is partially oxidised and split into 2 molecules of Pyruvic Acid (Pyruvate) Glycolysis occurs in the cytosol outside the mitochondrion. Glycolysis produces 2 molecules of ATP per glucose by substrate phosphorylation and 2 NADH.
2 2 NAD+ 2 NADH 2 H+

Glucose
2 ADP 2P

2 ATP

2 Pyruvate

Glycolysis Equation
Glycolysis consists of two coupled reactions - OXIDATION of GLUCOSE, an exergonic reaction
C6H12O6 + 2NAD+ ---> 2 NADH2 + 2 CH3COCOOH + ENERGY

-SYNTHESIS OF ATP an endergonic reaction 2 ATP + 2 H2O <-------- 2 ADP + 2 Pi + ENERGY There are 3 important products of glycolysis 2 Pyruvic acids which can enter the Krebs Cycle 2 NADH2 (NADH +H+) which can pass to the mitochondrion and generate 2 or 3 ATPs per NADH2 (<6 per glucose) 2 ATPs generated directly by glycolysis
Any reaction which forms ATP directly without using the electron transport chain is called

SUBSTRATE PHOSPHORYLATION

Substrate Phosphorylation
Enzyme A net yield of 2 ATP molecules are synthesised directly in GLYCOLYSIS P (without P using the electron transport chain) This is called P SUBSTRATE PHOSPHOR Phosphorylated YLATION. Organic substrate molecule (phosphate donor)

P ADP

Adenosine ATP

Phosphate transferred to ADP P

Steps of Glycolysis
Glucose STEPS IN GLYCOLYSIS When glucose first enters the cell, it is phosphorylated by ATP
A second phosphorylation occurs using a second ATP.
INTERSTITIAL FLUID CYTOSOL

Glucose-6-phosphate

The hexose is split into two trioses (glycerose), each of which continues in glycolysis Another phosphorylation occurs (using Pi) and NAD is reduced to NADH (2 NADH per glucose)
Each 3 C molecule is converted to pyruvic acid producing 2 ATP (4 ATP per glucose)

From mitochondria To mitochondria

1,3-Bisphosphoglyceric acid

ENERGY SUMMARY

Pyruvic acid
To mitochondria

Steps 1 & 2: 2 ATP Step 5: +4 ATP

NET GAIN: +2 ATP

Aerobic Glycolysis
2 NADH2 molecules result from GLYCOLYSIS of one GLUCOSE so a maximum of 6 ATP can be formed by
OXIDATIVE PHOSPHORYLATION (3 from each NADH). Since 2 ATP are formed directly during GLYCOLYSIS by SUBSTRATE PHOSPHORYLATION a maximum of 8 ATP can result from GLYCOLYSIS. Electron transport chain 6 ATP

2 Glucose

NAD+

2 NADH

2 H+

2 Pyruvate Substrate phosphorylation

2 ADP

2P

2 ATP

Anaerobic Glycolysis:Lactic Acid Fermentation

During vigorous exercise O2 is used up rapidly so anaerobic conditions occur in MUSCLE CELLS: pyruvate is reduced to Lactic Acid (lactate) instead of entering the mitochondrion. This process is called Lactic Acid Fermentation. Aerobically pyruvate enters the mitochondrion and is converted to acetyl coenzyme A which enters the Krebs Cycle.

Anaerobic Glycolysis:Lactic Acid Fermentation

2 NAD+

2 NADH

2 NADH

2 NAD+

GLYCOLYS IS 2 + 2 P ADP 2 ATP 2 Pyruvate 2 Lactate

Glucose

Anaerobically pyruvate is reduced to lactic acid (lactate) by NADH in Lactic Acid Fermentation Fermentation functions to regenerate NAD anaerobically so that glycolysis can continue (glycolysis needs NAD). The net energy yield from anaerobic GLYCOLYSIS is 2 ATP from substrate phosphorylation.

Formation of Acetyl Coenzyme A

Before entering the KREBS CYCLE PYRUVATE enters the MITOCHONDRION and is OXIDATIVELY DECARBOXYLATED, losing H2 and CO2.* H2 is taken by NAD forming NADH2 while the remaining ACETYL GROUP is combined with COENZYME A to form acetyl coenzyme A. COENZYME A is formed from the B vitamin PANTOTHENIC ACID and functions to deliver acetyl GROUPS to the KREBS CYCLE. Electron transport chain 6 ATP
NAD+ NADH

* Decarboxylations involve a coenzyme (cocarboxylase) containing vit B1, thyamine)

+ H+

CoA Pyruvate Acetyl CoA (acetyl coenzyme A)

CO2
Coenzyme A

The Krebs (TCA or Citric Acid) Cycle

The Krebs CYCLE is the final pathway for the CATABOLISM of various organic molecules in cells. The KREBS CYCLE takes 2 CARBON acetyl GROUPS and OXIDISES them by dehydrogenation forming CO2 as an end product. ACETYL GROUPS may come from pyruvate, fatty acids or from some amino acids. H2 from acetyl groups are fed into the electron transport chain by NAD or FAD forming ATP by oxidative phosphorylation.

Fatty acids

Glucose

Amino acids

Small carbon chains TCA cycle

Coenzymes

ATP Electron transport system

MITOCHONDRIA O2
CO2

H2O

The Krebs Cycle: Decarboxylations

The KREBS CYCLE operates only under aerobic conditions and occurs in the matrix of the MITOCHONDRION. The substrates of the Krebs cycle are various organic acids (referred to by their anion). ACETYL COENZYME A transfers the acetyl GROUP to the 4 carbon oxaloacetate to form the 6 carbon citrate (citric or tricarboxylic acid). 2 more DECARBOXYLATIONS occur. From each original GLUCOSE these reactions would form 4 CO2 molecules giving a total of 6 DECARBOXYLATIONS from each original GLUCOSE.

The Krebs Cycle: Sources of ATP

With each turn of the KREBS CYCLE from CITRATE to OXALOACETATE 3 NADH2 molecules are formed and a total of 9 ATP molecules will result from OXIDATIVE PHOSOPHORYLATION. These reactions generate 18 ATPs from each original GLUCOSE which entered GLYCOLYSIS. FAD acts as a H2 carrier in one case and this reaction will generate 2 ATPs from oxidative PHOSPHORYLATION (or 4 ATPs from each original GLUCOSE). One SUBSTRATE PHOSPHORYLATION occurs. The nucleotide GUANOSINE DIPHOSPHATE (GDP) is PHOSPHORYLATED to create a high energy molecule (GTP). GTP then transfers its phosphate to ADP to generate one ATP (2 from each glucose).

Pyruvic acid Coenzyme A CO2 Acetyl-CoA NADH Coenzyme A

Energy Yield from Krebs Cycle

4-carbon 2H

Citric acid 6-carbon CO2 2H 5-carbon CO2 2H NADH H2O NADH O2

ELECTRON TRANSPORT SYSTEM

2H

TCA CYCLE

ATP

4-carbon ATP FADH2 NADH

Copyright 2007 Pearson Education, Inc., publishing as Benjamin Cummings

Figure 17-4 7 of 7

Summary of Energy Yield from Krebs Cycle

Oxidative Decarboxylation of Pyruvate 3

Krebs Cycle

2 3 3

Summary of Cell Respiration

In cytoplasm

In cytoplasm

In mitochondrion

Gluconeogenesis
The liver has enzymes to reverse glycolysis, forming glucose from pyruvate or from oxaloacetate of the Krebs Cycle. This process is called GLUCONEOGENESIS (2) and converts lactic acid, glycerol & glucogenic amino acids to glucose - during exercise it converts LACTIC ACID back to glucose. which is sent back to the muscles to reenter glycolysis - when carbohydrates are unavailable as in starvation or diabetes mellitus gluconeogenesis maintains the blood glucose level (especially for brain cells). GLUCONEOGENESIS is stimulated by Glucocorticoids (mainly cortisol) released in times of stress via ACTH and by glucagon (both hyperglycemic hormones).

Functions of Glycogen
Carbohydrates can be stored in the body in the form of gycogen mainly in liver and muscle tissues.

Glycogenesis is the metabolic pathway by which

glycogen is formed from glucose.

Glycogenolysis is the metabolic pathway by which

glycogen is broken down to glucose or glucose - 6 - (P). The function of hepatic glycogenesis and glycogenolysis is to control the blood glucose level by storing glucose when blood sugar levels are high and releasing glucose when blood sugar levels are low. To release glucose to the blood cells must remove the phosphate from glucose 6 phosphate with the enzyme glucose 6 phosphatase. Liver cells have this enzyme but muscle cells do not. Muscle glycogen is primarily for use by muscle cells and muscle cells do not respond to glucagon.

Glycogenesis & Glycogenolysis

Glycogenesis Requires enzymes, glucokinase (or hexokinase) to phosphorylate glucose, glycogen synthetase (for 1-4 bonds), branching enzyme (for 1-6 bonds) in liver and skeletal muscle stimulated by insulin Glycogenolysis
enzymes phosphorylase (breaks 1-4 bonds), debranching enzyme (breaks 1-6 bonds)to form glucose 1-phosphate enzyme glucose 6 phosphatase releases glucose for the blood: only in hepatocytes so muscle cant release glucose Hepatic phosphorylase activated by glucagon & epinephrine, muscle glycogenolysis stimulated by epinephrine only.

Pathways in the Liver

Glycogen Glucose Hexokinase or Glucokinase Glucose-6-Pase Glucose-6-P Glucose + Pi Glycolysis Pathway

Glucose-1-P

Pyruvate Glucose metabolism in liver.

Glucose-6-phosphate may enter Glycolysis or (in liver) be dephosphorylated for release to the blood. Liver Glucose-6-phosphatase catalyzes the following, essential to the liver's role in maintaining blood glucose: glucose-6-phosphate + H2O glucose + Pi Most other tissues lack this enzyme.

Glycogen Storage Diseases

Glycogen Storage Diseases are Inborn Errors of Metabolism (enzyme deficiencies) associated with abnormal glycogen accumulation within liver or muscle cells. Some enzymes whose deficiency leads to glycogen accumulation are part of the inter-connected pathways shown here.

glycogen glucose-1-P Glucose-6-Phosphatase glucose-6-P glucose + Pi fructose-6-P Phosphofructokinase fructose-1,6-bisP Glycolysis continued

Effects of Glycogen Storage Diseases

When an enzyme defect affects mainly glycogen storage in liver, a common symptom is hypoglycemia, relating to impaired mobilization of glucose for release to the blood during fasting. When the defect is in muscle tissue, weakness & difficulty with exercise result from inability to increase glucose entry into Glycolysis during exercise. Additional symptoms depend on the particular enzyme that is deficient.

Examples of Glycogen Storage Diseases

Glycogen Storage Disease Type I, liver deficiency of Glucose-6-phosphatase (von Gierke's disease) Type IV, deficiency of branching enzyme in various organs, including liver (Andersen's disease) Type V, muscle deficiency of Glycogen Phosphorylase (McArdle's disease) Type VII, muscle deficiency of Phosphofructokinase. Symptoms, in addition to glycogen accumulation hypoglycemia (low blood glucose) when fasting, liver enlargement. liver dysfunction and early death.

muscle cramps with exercise.

inability to exercise.

Note: these glycogen storage diseases are autosomal recessive

Mitochondrial Disease
Mitochondrial disease is usually caused by an inherited defect in mitochondrial DNA. Mitochondria are inherited from the ova so are transmitted from mother to all offspring and from daughters to all their offspring. Mitochondrial disorders result in a lack of ATP in all cells causing: Lactic acidosis Fatigue Multiple organ damage, (e.g. brain, visual, auditory pathways).