TASK (P6/M4) a.

Describe using the biochemical pathway of glycolysis, the anaerobic breakdown of glucose to pyruvate Glucose is broken down in a series of steps, each catalysed by an enzyme. In the process, a small proportion of the energy in each glucose molecule is released, and used to make a small amount of ATP. The first steps in glycolysis involve adding phosphate groups to a glucose molecule. This produces a hexose sugar with two phosphate groups attached to it, a hexose bisphosphate. The process is called phosphorylation. It raises the energy level of the hexose, making it able to participate in the steps that follow. The hexose bisphosphate is then split into two molecules of a three-carbon sugar, triose phosphate. The trio phosphates are then oxidised to pyruvate, by having hydrogen removed from them. The enzyme that catalyses this reaction is called a dehydrogenase. It can only work if there is another molecule present that can take up the hydrogens that it removes.

b. Describe the fate of pyruvate under anaerobic conditions What happens to the pyruvate depends on the availability of oxygen in the cell. If there is plenty, then aerobic respiration can take place. The pyruvate is moved into a mitochondrion. This is done by active transport. Like a chloroplast, it is surrounded by an envelope of two membranes. The background material inside a mitochondrion is called the matrix. The link reaction Once inside the mitochondrion, the pyruvate undergoes a reaction known as the link reaction. This takes place in the matrix. During the link reaction, carbon dioxide is removed from the pyruvate. This is called decarboxylation, and it is catalysed by decarboxylase enzymes. The carbon dioxide is an excretory product, and it diffuses out of the mitochondrion and out of the cell. Pyruvate is a three-carbon substance, so the removal of carbon dioxide leaves a compound with two carbon atoms. At the same time as the carbon dioxide is removed, hydrogen is also removed from pyruvate. This is again picked up by NAD, producing reduced NAD. The reminder of the pyruvate combines with coenzyme A (often known as CoA) to produce acetyl CoA. The Krebs cycle The link reaction is given that name because it provides the link between the two main series of reactions in aerobic respiration-glycolysis and the Krebs cycle. The Krebs cycle takes place takes place in the matrix of the mitochondrion. It is a series of reactions in which a six-carbon compound is gradually changed to a four-carbon compound. First, the acetyl CoA made in the link reaction combines with a four-carbon compound called oxaloacetate. This converts oxaloacetate into a six-carbon compound called citrate. In a series of small steps, the citrate is converted back to oxaloacetate. As this happens, more carbon dioxide is released and more NAD is reduced as it accepts hydrogen. In one stage, a different coenzyme, called FAD, accepts hydrogen. And at one point in the cycle a molecule of ATP is made. Each of the steps in the Krebs cycle is catalysed by a specific enzyme. These enzymes are all present in the matrix of the mitochondrion. Those that cause oxidation are called oxidoreductases or dehydrogenases. Those that remove carbon dioxide are decarboxylases.

Oxidative phosphorylation The last stages of aerobic respiration involve oxidative phosphorylation: the use of oxygen to produce ATP from ADP and Pi. The electron transport chain Held in the inner membrane of the mitochondrion are molecules called electron carriers. They make up the electron transport chain. Each reduced NAD molecule- which was reduced in the matrix during the Krebs cyclereleases its hydrogens. Each hydrogen atom splits into a hydrogen ion, H+ (a proton) and an electron-. H H+ + e-

The electrons are picked up by the first of the electron carriers. The carrier is now reduced, because it has gained an electron. The reduced NAD has been oxidised, because it has lost hydrogen. The NAD can now go back to the Krebs cycle and be re-used as a coenzyme to pick up hydrogen. The first electron carrier passes its electron to the next in the chain. The first carrier is therefore oxidised (because it has lost an electron) and the second is reduced. The electron is passed from one carrier to the next all the way along the chain. As the electron is moved along, it releases energy which is used to make ATP. At the end of the electron transport chain, the electron combines with a hydrogen ion and with oxygen, to form water. This is why we need oxygen. The oxygen acts as the final electron acceptor for the electron transport train.

TASK (D4) Explain how glycolysis is regulated to meet the energy demands of the cell There are ten enzymes that catalyse the steps in glycolysis that convert glucose into pyruvate, and the entire pathway is located in the cytoplasm of eukaryotic cells. The activity of the pathway is regulated at key steps to ensure that glucose consumption and energy production match the needs of the cell. The steps along the pathway each involve a change in the free energy of the products and reactants, and as long as the overall change in free energy is negative, the reaction continues forward, like water flowing down hill to its lowest energy point. The key steps in the regulation of glycolysis, or any pathway, are those that catalyse the rate-limiting, irreversible steps along the pathway. In glycolysis in mammals, the key regulatory enzyme is phosphofructokinase, which catalyses the rate-limiting committed step. Phosphofructokinase is activated by AMP and inhibited by ATP, among other regulatory mechanisms. Thus, when ATP is low (and AMP is high), phosphofructokinase will be activated and generate more ATP. Similarly, when ATP is abundant, phosphofructokinase will be inhibited to prevent wasting glucose on making energy when it is not needed. Although the glycolytic pathway was one of the first studied, it is still relevant to many issues faced in modern biology. Failure to provide energy can have lethal consequences for cells - the absence of oxygen caused by a stroke or a heart attack that prevents ATP generation can have lethal consequences for the cells involved. Cancer cells often generate energy through glycolytic fermentation more than oxidative phosphorylation, suggesting that manipulation of metabolism may provide a therapeutic strategy. Well known glycolytic enzymes such as glyceraldehyde-3-phosphate dehydrogenase may play roles in other cellular processes such as apoptosis. Future studies may reveal additional functions of glycolytic enzymes.

Gluconeogenesis and glycolysis are coordinated so that within a cell one pathway is relatively inactive while the other is highly active. If both sets of reactions were highly active at the same time, the net result would be the hydrolysis of four nucleotide triphosphates (two ATP plus two GTP) per reaction cycle. Both glycolysis and gluconeogenesis are highly exergonic under cellular conditions, and so there is no thermodynamic barrier to such simultaneous activity. However, the amounts and activities of the distinctive enzymes of each pathway are controlled so that both pathways are not highly active at the same time. The rate of glycolysis is also determined by the concentration of glucose and the rate of gluconeogenesis by the concentrations of lactate and other precursors of glucose.

The glycolytic pathway consists of 10 enzyme-catalyzed steps. During glycolysis, glucose, a six-carbon carbohydrate, is oxidized to form two molecules of pyruvate, a three-carbon molecule. For each glucose molecule metabolized, the pathway produces two molecules of ATP and two molecules of NADH. Glycolysis is not isolated from other metabolic pathways. Other molecules besides glucose can enter at a few points along the glycolytic pathway. For example, the product of glycogen breakdown, glucose-6-phosphate, can enter the glycolytic pathway at the second step. Glyceraldehyde-3-phosphate, which is produced by photosynthesis, is also a glycolytic intermediate, so it can be directed from this anabolic pathway into glycolysis when energy is needed. Additionally, intermediates can be drawn out of the glycolytic pathway when energy levels are high, for use in biosynthetic pathways. For instance, during active energy production pyruvate, the product of glycolysis, enters the citric acid cycle, but when energy is not needed pyruvate serves as a substrate in amino acid synthesis. The pyruvate produced has one of three metabolic fates, to either become acetyl-CoA, ethanol, or lactate. http://www.biocarta.com/pathfiles/h_glycolysispathway.asp

http://biochemistrystudy.blogspot.co.uk/2012/10/fates-of-pyruvate-under-aerobicand.html

http://themedicalbiochemistrypage.org/glycolysis.php

The pyruvate produced has one of three metabolic fates, to either become acetyl-CoA, ethanol, or lactate.

http://www.biocarta.com/pathfiles/h_glycolysispathway.asp http://www.wiley.com/college/pratt/0471393878/student/structure/glycolysis/

Glycolysis is the almost universal pathway that converts glucose into pyruvate. In aerobic organisms the pyruvate passes into the mitochondria where it is completely oxidised by O2 into CO2 and H2O and its potential energy largely conserved as ATP. In the absence of sufficient oxygen the pyruvate is reduced by the NADH to a wide range of products, especially lactate in animals and ethanol in yeasts. The glycolytic pathway is highly unregulated in rapidly-growing malignant tumour cells, a phenomenon first described by Ottow Warburg in 1930. This phenomenon, commonly referred to as the Warburg effect, is a preference for highly proliferative active cells to shift to aerobic glycolysis even in the presence of adequate oxygen. In aerobic glycolysis, NADH is regenerated through the reduction of pyruvate to lactic acid by lactate dehydrogenase. Enzymes of the glycolytic pathway are potential therapeutic targets for the treatment of cancer. Under anaerobic conditions (as in very active skeletal muscles, in submerged plants, or in lactic acid bacteria, for example), NADH generated by glycolysis cannot be deoxidised by oxygen. Failure to regenerate NAD+ would leave the cell with no electron acceptor for the oxidation of glyceraldehyde-3-phosphate, and the energy-yielding reactions of glycolysis would stop. NAD+ must therefore be regenerated by some other reaction. The earliest cells to arise during evolution lived in an atmosphere almost devoid of oxygen and had to develop strategies for carrying out glycolysis under anaerobic conditions. Most modern organisms have retained the ability to continually regenerate NAD+ during anaerobic glycolysis by transferring electrons from NADH to form a reduced end product such as lactate or ethanol.

In glycolysis, the six-carbon sugar glucose is oxidized and split in two halves, to create two molecules of pyruvate (3 carbons each) from each molecule of glucose. Along the way, the cell extracts a relatively small amount of energy from glucose in the form of ATP, 2 ATP molecules collected for each glucose molecule that starts down the glycolytic path. The pyruvate produced has one of three metabolic fates, to either become acetyl-CoA, ethanol, or lactate. When oxygen is available, the pyruvate can be converted to acetyl-CoA and enter the Krebs cycle, where the acetyl-CoA will be completely oxidized and generate ATP through oxidative phosphorylation. Fermentation is much less efficient than oxidative phosphorylation in making ATP, creating only 2 ATP per glucose while oxidative phosphorylation creates 36 ATP per glucose in mammalian cells. Oxidative phosphorylation does not work in the absence of oxygen, however, and in the absence of oxygen glycolysis is forced to a halt due to a lack of NAD+, unless NAD+ is regenerated through fermentation. In yeast, fermentation allows the yeast to continue producing energy and survive in the absence of oxygen, producing ethanol and carbon dioxide from pyruvate. In mammalian muscle, strenuous exertion can create conditions in which oxygen is consumed faster than blood can provide it, forcing the muscle to use fermentation and create lactic acid in this case and make your muscles sore after a workout.