AP Biology John D. O’Bryant School of Mathematics and Science October 11, 2012 AP Biology Agenda Do Now (Quiz) Experimental Design: Lactose Intolerance, etc. AP Biology Do Now (Quiz) 1. Besides turning enzymes on or off, what other means does a cell use to control enzymatic activity in the cell? A) cessation of all enzyme formation B) compartmentalization of enzymes into defined organelles C) exporting enzymes out of the cell D) connecting enzymes into large aggregates E) hydrophobic interactions AP Biology Do Now (Quiz) 2. Which of the following is an example of cooperativity? A) the binding of an end product of a metabolic pathway to the first enzyme that acts in the pathway B) protein function at one site affected by binding at another of its active sites C) a molecule binding at one unit of a tetramer allowing faster binding at each of the other three D) the effect of increasing temperature on the rate of an enzymatic reaction E) binding of an ATP molecule along with one of the substrate molecules in an active site AP Biology Do Now (Quiz) 3. Which of the following is the most correct interpretation of the figure? A) Inorganic phosphate is created from organic phosphate. B) Energy from catabolism can be used directly for performing cellular work. C) ADP + Pi are a set of molecules that store energy for catabolism. D) ATP is a molecule that acts as an intermediary to store energy for cellular work. E) Pi acts as a shuttle molecule to move energy from ATP to ADP. AP Biology Do Now (Quiz) 4. How does a noncompetitive inhibitor decrease the rate of an enzyme reaction? A) by binding at the active site of the enzyme B) by changing the shape of a reactant C) by changing the free energy change of the reaction D) by acting as a coenzyme for the reaction E) by decreasing the activation energy of the reaction AP Biology Do Now (Quiz) 5. Increasing the substrate concentration in an enzymatic reaction could overcome which of the following? A) denaturization of the enzyme B) allosteric inhibition C) competitive inhibition D) saturation of the enzyme activity E) insufficient cofactors AP Biology Metabolism & Enzymes AP Biology 2007-2008 Factors that Affect Enzymes AP Biology 2007-2008 Factors Affecting Enzyme Function Enzyme concentration Substrate concentration Temperature pH Salinity Activators Inhibitors AP Biology catalase Enzyme concentration reaction rate What’s happening here?! enzyme concentration AP Biology Factors affecting enzyme function Enzyme concentration as enzyme = reaction rate more enzymes = more frequently collide with substrate reaction rate levels off reaction rate substrate becomes limiting factor not all enzyme molecules can find substrate AP Biology enzyme concentration Substrate concentration reaction rate What’s happening here?! substrate concentration AP Biology Factors affecting enzyme function Substrate concentration as substrate = reaction rate more substrate = more frequently collide with enzyme reaction rate levels off reaction rate all enzymes have active site engaged enzyme is saturated maximum rate of reaction AP Biology substrate concentration Temperature reaction rate What’s happening here?! 37° temperature AP Biology 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 AP Biology Enzymes and temperature Different enzymes function in different organisms in different environments reaction rate human enzyme hot spring bacteria enzyme 37°C AP Biology temperature 70°C (158°F) How do ectotherms do it? AP Biology pH What’s happening here?! trypsin reaction rate pepsin pepsin trypsin 0 AP Biology 1 2 3 4 5 6 pH 7 8 9 10 11 12 13 14 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 (end 10/11) optimal pH? most human enzymes = pH 6-8 depends on localized conditions pepsin (stomach) = pH 2-3 trypsin (small intestines) = pH 8 AP Biology 0 1 2 3 4 5 6 7 8 9 10 11 Salinity reaction rate What’s happening here?! salt concentration AP Biology 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! AP Biology Compounds which help enzymes Fe in Activators hemoglobin cofactors non-protein, small inorganic compounds & ions Mg, K, Ca, Zn, Fe, Cu bound within enzyme molecule coenzymes non-protein, organic molecules bind temporarily or permanently to enzyme near active site AP Biology many vitamins NAD (niacin; B3) FAD (riboflavin; B2) Coenzyme A Mg in chlorophyll Compounds which regulate enzymes Inhibitors molecules that reduce enzyme activity competitive inhibition noncompetitive inhibition irreversible inhibition feedback inhibition AP Biology Competitive Inhibitor Inhibitor & substrate “compete” for active site penicillin blocks enzyme bacteria use to build cell walls disulfiram (Antabuse) treats chronic alcoholism blocks enzyme that breaks down alcohol severe hangover & vomiting 5-10 minutes after drinking Overcome by increasing substrate concentration AP Biology saturate solution with substrate so it out-competes inhibitor for active site on enzyme Non-Competitive Inhibitor Inhibitor binds to site other than active site allosteric inhibitor binds to allosteric site causes enzyme to change shape conformational change active site is no longer functional binding site keeps enzyme inactive some anti-cancer drugs inhibit enzymes involved in DNA synthesis stop DNA production stop division of more cancer cells cyanide poisoning irreversible inhibitor of Cytochrome C, an enzyme in cellular respiration stops production of ATP AP Biology Irreversible inhibition Inhibitor permanently binds to enzyme competitor permanently binds to active site allosteric permanently binds to allosteric site permanently changes shape of enzyme nerve gas, sarin, many insecticides (malathion, parathion…) cholinesterase inhibitors AP Biology doesn’t breakdown the neurotransmitter, acetylcholine Allosteric regulation Conformational changes by regulatory molecules inhibitors keeps enzyme in inactive form activators keeps enzyme in active form AP Biology Conformational changes Allosteric regulation Metabolic pathways ABCDEFG 5 6 enzyme enzyme enzyme enzyme enzyme enzyme enzyme 1 2 3 4 Chemical reactions of life are organized in pathways AP Biology divide chemical reaction into many small steps artifact of evolution efficiency intermediate branching points control = regulation Efficiency Organized groups of enzymes enzymes are embedded in membrane and arranged sequentially Link endergonic & exergonic reactions Whoa! All that going on in those little mitochondria! AP Biology Feedback Inhibition Regulation & coordination of production product is used by next step in pathway final product is inhibitor of earlier step allosteric inhibitor of earlier enzyme feedback inhibition no unnecessary accumulation of product ABCDEFG 1 2 3 4 5 6 X enzyme enzyme enzyme enzyme enzyme enzyme AP Biology allosteric inhibitor of enzyme 1 threonine Feedback inhibition Example synthesis of amino acid, isoleucine from amino acid, threonine isoleucine becomes the allosteric inhibitor of the first step in the pathway as product accumulates it collides with enzyme more often than substrate does AP Biology isoleucin e Don’t be inhibited! Ask Questions! AP Biology 2007-2008 Cooperativity Substrate acts as an activator substrate causes conformational change in enzyme induced fit favors binding of substrate at 2nd site makes enzyme more active & effective hemoglobin Hemoglobin 4 polypeptide chains can bind 4 O2; 1st O2 binds now easier for other O2 to bind AP3Biology Lorenzo’s Oil AP Biology