2. Ch.8 An Introduction to Metabolism

3. Flow of energy through life Life is built on chemical reactions –transforming energy from one form to another organic molecules  ATP & organic molecules sun solar energy  ATP & organic molecules

4. Metabolism –Is the totality of an organism’s chemical reactions

5. Organization of the Chemistry of Life into Metabolic Pathways A metabolic pathway begins with a specific molecule and ends with a product Each step is catalyzed by a specific enzyme Starting molecule Product Enzyme 1 Enzyme 2 Enzyme 3 B C A D

6. Metabolism Chemical reactions of life –forming bonds between molecules dehydration synthesis synthesis anabolic reactions –breaking bonds between molecules hydrolysis digestion catabolic reactions That’s why they’re called anabolic steroids!

7. Thermodynamics Energy (E)~ capacity to do work; Kinetic energy~ energy of motion; Potential energy~ stored energy Thermodynamics~ study of E transformations 1st Law: conservation of energy; E transferred/transformed, not created/destroyed 2nd Law: transformations increase entropy (disorder, randomness)

8. Free energy Free energy: portion of system’s E that can perform work (at a constant T) Exergonic reaction: net release of free E to surroundings Endergonic reaction: absorbs free E from surroundings

9. Change in free energy, ∆G The change in free energy, ∆G during a biological process –Is related directly to the enthalpy change (∆H) and the change in entropy (∆S) ∆G = ∆H – T∆S T = temp in Kelvin (K)

10. Chemical reactions & energy Some chemical reactions release energy –exergonic –∆G < 0 –spontaneous –digesting polymers –hydrolysis = catabolism digesting molecules= LESS organization= lower energy state Figure 8.6 Reactants Products Energy Progress of the reaction Amount of energy released (∆G <0) Free energy (a) Exergonic reaction: energy released

11. Chemical reactions & energy Some chemical reactions require input of energy –endergonic –∆G > 0 –non-spontaneous –building polymers –dehydration synthesis = anabolism building molecules= MORE organization= higher energy state Figure 8.6 Energy Products Amount of energy released (∆G>0) Reactants Progress of the reaction Free energy (b) Endergonic reaction: energy required

12. The energy needs of life Organisms are endergonic systems –What do we need energy for? synthesis –building biomolecules reproduction movement active transport temperature regulation

13. Where do we get the energy from? Work of life is done by energy coupling –use exergonic (catabolic) reactions to fuel endergonic (anabolic) reactions ++ energy + + digestion synthesis

14. ATP Living economy Fueling the body’s economy –eat high energy organic molecules food = carbohydrates, lipids, proteins, nucleic acids –break them down digest = catabolism –capture released energy in a form the cell can use Need an energy currency –a way to pass energy around –need a short term energy storage molecule Whoa! Hot stuff!

15. ATP high energy bonds How efficient! Build once, use many ways Adenosine TriPhosphate –modified nucleotide nucleotide = adenine + ribose + P i  AMP AMP + P i  ADP ADP + P i  ATP –adding phosphates is endergonic

16. How does ATP store energy? P O–O– O–O– O –O–O P O–O– O–O– O –O–O P O–O– O–O– O –O–O P O–O– O–O– O –O–O P O–O– O–O– O –O–O P O–O– O–O– O –O–O P O–O– O–O– O –O–O P O–O– O–O– O –O–O Each negative PO 4 more difficult to add –a lot of stored energy in each bond most energy stored in 3rd P i 3rd P i is hardest group to keep bonded to molecule Bonding of negative P i groups is unstable –spring-loaded –P i groups “pop” off easily & release energy Instability of its P bonds makes ATP an excellent energy donor I think he’s a bit unstable… don’t you? AMP ADPATP

17. How does ATP transfer energy? P O–O– O–O– O –O–O P O–O– O–O– O –O–O P O–O– O–O– O –O–O 7.3 energy + P O–O– O–O– O –O–O ATP  ADP –releases energy ∆G = -7.3 kcal/mole Fuel other reactions Phosphorylation –released P i can transfer to other molecules destabilizing the other molecules –enzyme that phosphorylates = “kinase” ADPATP

18. It’s never that simple! An example of Phosphorylation… Building polymers from monomers –need to destabilize the monomers –phosphorylate! C H OH H HOHO C C H O H C + H2OH2O + +4.2 kcal/mol C H OH C H P + ATP + ADP H HOHO C + C H O H CC H P + PiPi “kinase” enzyme -7.3 kcal/mol -3.1 kcal/mol enzyme H OH C H HOHO C synthesis

19. Can’t store ATP  good energy donor, not good energy storage too reactive transfers P i too easily only short term energy storage  carbohydrates & fats are long term energy storage ATP / ADP cycle A working muscle recycles over 10 million ATPs per second Whoa! Pass me the glucose (and O 2 )! ATP ADP PiPi + 7.3 kcal/mole cellular respiration

20. Another example of Phosphorylation… The first steps of cellular respiration –beginning the breakdown of glucose to make ATP glucose C-C-C-C-C-C fructose-1,6bP P-C-C-C-C-C-C-P DHAP P-C-C-C G3P C-C-C-P hexokinase phosphofructokinase Those phosphates sure make it uncomfortable around here! C H P C P C ATP 2 ADP 2 activation energy

21. Too much activation energy for life Activation energy –amount of energy needed to destabilize the bonds of a molecule –moves the reaction over an “energy hill” glucose

22. Reducing Activation energy Catalysts –reducing the amount of energy to start a reaction Pheeew… that takes a lot less energy! reactant product uncatalyzed reaction catalyzed reaction NEW activation energy

23. Catalysts So what’s a cell got to do to reduce activation energy? –get help! … chemical help… ENZYMES GG Call in the ENZYMES!

24. Substrate Specificity of Enzymes The substrate –Is the reactant an enzyme acts on The enzyme –Binds to its substrate, forming an enzyme-substrate complex The active site –Is the region on the enzyme where the substrate binds Figure 8.16 Substrate Active site Enzyme (a)

25. Naming conventions Enzymes named for reaction they catalyze –sucrase breaks down sucrose –proteases break down proteins –lipases break down lipids –DNA polymerase builds DNA adds nucleotides to DNA strand –pepsin breaks down proteins (polypeptides)

26. Factors Affecting Enzyme Function Enzyme concentration Substrate concentration Temperature pH Salinity Activators Inhibitors catalase

27. Factors affecting enzyme function Enzyme concentration –as  enzyme =  reaction rate more enzymes = more frequently collide with substrate –reaction rate levels off substrate becomes limiting factor not all enzyme molecules can find substrate enzyme concentration reaction rate

28. Factors affecting enzyme function substrate concentration reaction rate Substrate concentration –as  substrate =  reaction rate more substrate = more frequently collide with enzyme –reaction rate levels off all enzymes have active site engaged enzyme is saturated maximum rate of reaction

29. Factors affecting enzyme function Temperature –Optimum T° greatest number of molecular collisions human enzymes = 35°- 40°C –body temp = 37°C –Heat: increase beyond optimum T° increased energy level of molecules disrupts bonds in enzyme & between enzyme & substrate –H, ionic = weak bonds denaturation = lose 3D shape (3° structure) –Cold: decrease T° molecules move slower decrease collisions between enzyme & substrate

30. Enzymes and temperature Different enzymes function in different organisms in different environments 37°C temperature reaction rate 70°C human enzyme hot spring bacteria enzyme (158°F)

31. Factors affecting enzyme function pH –changes in pH adds or remove H + disrupts bonds, disrupts 3D shape –disrupts attractions between charged amino acids –affect 2° & 3° structure –denatures protein –optimal pH? most human enzymes = pH 6-8 –depends on localized conditions –pepsin (stomach) = pH 2-3 –trypsin (small intestines) = pH 8 72013456891011

32. Factors affecting enzyme function Salt concentration –changes in salinity adds or removes cations (+) & anions (–) disrupts bonds, disrupts 3D shape –disrupts attractions between charged amino acids –affect 2° & 3° structure –denatures protein –enzymes intolerant of extreme salinity Dead Sea is called dead for a reason!

33. Enzyme cofactors Cofactors –Are non-protein enzyme helpers e.g. zinc, iron, copper atoms Coenzymes –Are organic cofactors e.g. vitamins

34. coenzymes in group transfer reactions coenzyme abbreviation entity transfered nicotine adenine dinucelotide NAD - partly composed of niacin electron (hydrogen atom) nicotine adenine dinucelotide phosphate NADP -Partly composed of niacin electron (hydrogen atom) flavine adenine dinucelotide FAD - Partly composed of riboflavin (vit. B2) electron (hydrogen atom) coenzyme A CoA Acyl groups coenzymeQ CoQ electrons (hydrogen atom)

35. Enzyme Regulation Regulation of enzyme activity helps control metabolism A cell’s metabolic pathways –Must be tightly regulated

36. Enzyme Inhibitors Competitive inhibitors –Bind to the active site of an enzyme, competing with the substrate Figure 8.19 (b) Competitive inhibition A competitive inhibitor mimics the substrate, competing for the active site. Competitive inhibitor A substrate can bind normally to the active site of an enzyme. Substrate Active site Enzyme (a) Normal binding

37. Enzyme Inhibitors Noncompetitive inhibitors –Bind to another part of an enzyme, changing the function Figure 8.19 A noncompetitive inhibitor binds to the enzyme away from the active site, altering the conformation of the enzyme so that its active site no longer functions. Noncompetitive inhibitor (c) Noncompetitive inhibition

38. Allosteric regulation Conformational changes by regulatory molecules –inhibitors keeps enzyme in inactive form –activators keeps enzyme in active form Conformational changesAllosteric regulation

39. Feedback inhibition –The end product of a metabolic pathway shuts down the pathway Active site available Isoleucine used up by cell Feedback inhibition Isoleucine binds to allosteric site Active site of enzyme 1 no longer binds threonine; pathway is switched off Initial substrate (threonine) Threonine in active site Enzyme 1 (threonine deaminase) Intermediate A Intermediate B Intermediate C Intermediate D Enzyme 2 Enzyme 3 Enzyme 4 Enzyme 5 End product (isoleucine) Figure 8.21

40. Cooperativity Substrate acts as an activator –substrate causes conformational change in enzyme induced fit –favors binding of substrate at 2 nd site –makes enzyme more active & effective hemoglobin Hemoglobin  4 polypeptide chains  can bind 4 O 2 ;  1 st O 2 binds  now easier for other 3 O 2 to bind