10/10/05 2:45 PM

Controls of the Glycolytic pathway

Very sensitive to the level of G6P in the cell. The enzyme is inhibited by increases in the level of G6P to prevent excess trapping of G6P. Trapping results from adding a phosphate to the glucose molecule. This trapping process requires one ATP to be reduced to ADP. Phosphofructokinase-1 (PFK-1) Responsible for converting Fructose 6 Phosphate (F6P) into Fructose 1,6 Bisphosphate (F1,6BisP)

The switch that controls glycolysis. PFK-1 is activated by increased levels of AMP and inhibited when levels become low.

Aside: Where does AMP come from? When ATP is used in the cell ADP is a resulting product. When there is an excess of ADP in the cell Adenylate kinase is activated. Adenylate kinase performs a very simple reversible reaction. 2 ADP AMP + ATP When there is an increase in AMP levels, adenylate kinase begins to move in the reverse direction and communicates to PFK-1 that there is excess ADP. There is a continual eb and flow between ATP usage, PFK-1 activation and ATP formation. Pyruvate Kinase Responsible for converting Enolpyruvate into pyruvate.

This recreation is affected by fructose 1-6 Bisphosphate (+ve) and is inhibited by covalent modification (-ve).

When there is excess inorganic phosphate (Pi) in the system it binds with pyruvate kinase and inhibits its action. Glycolysis in the Liver The main function of the liver is to maintain glycogen homeostasis. The liver uses glucokinase to trap (add a phosphate) glucose it is not inhibited by product (glucose 6 phosphate) so it will keep trapping

Glucose regulation in the liver The liver will maintain a concentration of 4-6 Mmol/L of glucose in the blood. After a meal these levels can increase to 7 Mmol/L of blood. Excess glucose (i.e. after a meal) the liver will convert glucose into glycogen. This stored glycogen can be used when the body is in the fasting state. Starved state (i.e. skipped a meal) the liver will break stored glycogen into glucose and release it into the blood to maintain a concentration of 4-6 Mmol/L. Excess glycogen When glycogen storage space is filled the liver will begin storing excess glucose as fat. To complete this process glucose must be coverted to pyruvate. Due to the high levels of ATP in the liver, glycolysis should not occur because ATP inhibits PKF1 (rate controlling enzyme). Hormonal regulation Insulin is released after a meal to indicate the fed state. It activates Glucokinase, PFK1 and pyruvate kinase to encourage substrate storage (as glycogen or fat) Glucagon is released in the fasting state. This inhibits glucokinase (stop traping glucose), PFK1 and pyruvate kinase (stop storing substrates). Fructose 2,6-Bisphosphate (f2,6BisP) This molecule overrides the inhibitory effects of ATP on PFK1 and signals the liver cells to convert glucose 6-phosphate to pyruvate. It is not a part of the pathway, but merely a signal. It is created by and reverted to fructose 6-Phosphate. There is excess Fructose 6-phosphate when glycogen stores are filled. This activates PFK2 to convert F6P to fructose 2,6 Bisphosphate (F2,6BisP)

(Remember: this occurs in liver cells in the fed state)

More on PFK2 PFK2 is referred to as an enzyme in tandem because it is actually two enzymes on one chain. It contains both the PFK2 and fructose 2-6 Bisphosphate. PFK2 is active in the fed state (insulin is present) and converts F6P to F2,6BisP to activate PFK1. F2,6BisPase is active in the fasting state (glucagon is present) and reverts F2,6BisP to F6P. Protein kinase allows a phosphate to bind to the enzyme to activate Fructose 2,6 Bisphophatase and protein phosphatase removes the phosphate to activate PFK2.

Non rate controlling enzymes Glyceraldehyde 3-Phosphate dehydrogenase Converts Glyceraldehyde 3 phosphate (Ga3P) into 1,3-Bisphosphoglycerate

If Ga3P is exposed to arsenic, it will still move through the reaction but arsenic will replace phosphate in 1,3 Bisphosphoglycerate to create 1 arseno 3 phosphoglycerate. Glycolysis will continue except when 1 arseno 3 phosphoglycerate reaches the stage where a phosphate is removed to produce ATP the arsenic is removed instead result in zero net ATP production. This can effect ATP production is many tissues, including nervous tissue and lead to cell damage or death. Enolase: Converts 2-Phosphoglycerate to Phosphoenologlycerate

Effects of fluoride Fluoride (higher affinity) removes Mg2+ from enolase and stops the reaction. This process is called chelation. This is used in the process of blood-glucose testing when the blood cannot be tested directly after removal. Fluoride is added to the blood so the sample stops processing glucose and accurate measures can be taken. The level of fluoride in your toothpaste or water cannot causes these affects in the body.

Red blood cell uses Na+/K+ atpase This concentration gradient helps keep the cell shape. o removes 3 sodium (Na+) ions and imports 2 potassium (K+) in some cells this requires up to 40% of atp to maintain balance o This process requires 1 ATP + 1 H20 and produces 1 ADP and 1 Pi The red blood cell also has a special enzyme: bisphosphoglycerate mutase and produces 2,3-biosphophoglycerate

o this molecule is required for proper hemoglobin activity. o It binds to hemoglobin and stabilizes hemoglobin in the deoxy-form (without oxygen). This accounts for how hemoglobin reacts physiologically. o Originally though to be a specific waste product. But then it was found that oxygen would not bind the same way when it was not present. It can be broken down by Bisphosphoglycerate mutase Excess lactate can lower the pH of a cell and do damage, red blood cells will release this into the blood. Tissues that support anaerobic glycolysis RBC, WBC kidney medulla skin tissues of the eye skeletal muscle

The Cori Cycle A cell (i.e. RBC) uses a substrate (glucose) in anaerobic glycolysis to produce energy and releases its waste product (lactate) into the blood stream. This is then used in the liver as a substrate for glucogenesis to create a product (glucose). Other carbohydrates For next Tuesday 1. Describe the metabolism of fructose in liver and other tissues (chpt 12) 2. Describe the metabolism of galactose in the liver (12) 3. Compare and contrast two types of galactosemia (12) 4. Describe the constituents of the extra cellular matrix (4,14) From biochemistry Harvey and Champe (2005) The main theme is convert the CHO into a suitable substrate for the glycolytic pathway. Mannose Use hexokinase to convert to mannose 6P then use phosphomannose isomerase to create fructose 6-P. since phosphomannose isomerase is reversible you can make mannose 6-P or break it down depending on the content in the diet and the bodies requirements.

Mannose is required for the formation of glycoproteins. If there is insufficient mannose in the diet, glucose is converted G6P, then fructose 6 phosphate and that is converted to mannose 6P using phophomannose isomerase

Fructose supplies 10% (50g) of the daily calories (i.e sugar in coffee) Does not stimulate pancreas to secrete insulin (will not cause fedstate) found in fruits and honey I like honey too Entry into all tissues is independent of insulin Certain sugar have higher sweetness than other. Fructose has the highest, followed by invert sugar (glucose + fructose). Metabolism o Using hexokinase to make fructose 6P, but it requires an abnormally high concentration of fructose o The liver receives fructose first and traps it with a P to make fructose 1P with fructokinase.

it then uses aldelase B to cut it in two: glyceraldehyde and dihydroacetone phosphate

o However, since glyceraldehyde contains no P, it is not trapped.

So trikinase is used to add a P and trap it.

o Glyceraldehyde can be reduced with alcohol dehydrogenase to form glycerol.

o fructose can be converted to fat more rapidly than glucose. glucose is made into glycogen first, and is dependant on PFK1. Fructose bypasses PFK1 (which is the rate limiting step).

Essential fructosuria (inborn error) lack fructose kinase and the liver cant trap fructose. It is lost into the system, pass the renal threshold for fructose and appear in the urine. The solution to this is to remove fructose and sucrose from the diet.

Fructose intolerance unable to convert past fructose 1-P due to a problem with aldelase B. Excess fructose can eventually destroy the cell. This could lead to liver failure, and problems at the kidney and small intestine. The solution again is to remove fructose from the diet.