October 11 AP Biology

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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

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




ABCDEFG
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



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

ABCDEFG
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
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