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They make things, they break things
Enzymes They make things, they break things
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Enzymes: Function Inhibition Product Inhibition Activators Cofactors
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What is an Enzyme? Most enzymes are globular proteins
Definition: “A macromolecule serving as a catalyst, a chemical agent that changes the rate of a reaction without being consumed by the reaction”. An RNase
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Simulation: Observing an enzyme in action
18 volunteers (2 from each group) Chemical reaction simulation
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Simulation: Test Reaction:
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Method:
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Enzyme Observations:
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Enzymes: Not used up in the reaction.
Make the reaction require less energy. One enzyme molecule can be a catalyst for several reactions. Usually end in the suffix “-ase”. E.g. DNA polymerase, ATP synthase, Ligase, Sucrase, RNase, etc. The FUNCTION of an enzyme is often indicated by the enzyme name.
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Rate of Reaction: What would happen to the rate of reaction if more substrate was added? What would happen to the rate of reaction if more enzyme was added?
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Why Enzymes? Energy required for RXN to occur.
Energy required for RXN to occur IF enzymes used. HEAT
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Why Enzymes? Enzymes lower the required Energy of Activation
This allows for faster reactions This facilitates reactions that otherwise could not occur as quickly Some enzymes can make reactions go trillions of times faster Carbonic anhydrase (stomach) can catalyze over 1,000,000 reactions per second Some enzymes are temperature/pH specific.
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A B C D Transition state A B EA C D Free energy Substrate A B ∆G < O C D Products Progress of the reaction
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Progress of the reaction
Course of reaction without enzyme EA without enzyme EA with enzyme is lower Substrate Free energy ∆G is unaffected by enzyme Course of reaction with enzyme Products Progress of the reaction
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How Do Enzymes Work?
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How Do Enzymes Work? Orienteer: Stressor: Good microenvironment:
Active site encourages substrates to come together in the correct orientation Stressor: Induced fit stresses and bends critical chemical bonds. Good microenvironment: E.g. Active site is low pH. Direct contribution: Brief covalent bonds between substrate and R groups.
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Substrate Active site Enzyme Enzyme-substrate complex (a) (b)
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Sucrose + H2O + Sucrase Sucrase-Sucrose-H2O Complex Glucose + Fructose + Sucrase
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Substrates enter active site; enzyme
changes shape such that its active site enfolds the substrates (induced fit). 1 Substrates held in active site by weak interactions, such as hydrogen bonds and ionic bonds. 2 Substrate Enzyme-substrate complex Active site can lower EA and speed up a reaction. 3 Active site is available for two new substrate molecules. 6 Enzyme 5 Products are released. Substrates are converted to products. 4 Products
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Specificity Enzymes often catalyze only a single reaction
Enzymes are typically very specific How many different kinds of enzymes are in the human body?
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Enzymes: Function Inhibition Product Inhibition Activators Cofactors
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Minute Paper What are the top three things you learned about enzyme function?
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Enzymes: Function Inhibition Product Inhibition Activators Cofactors
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Enzyme Inhibition Enzymes can be inhibited by compounds
Inhibitor often similar in structure to substrate Enzymes can be inhibited at active site or a different part of the molecule Allosteric inhibition
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Enzyme Inhibition Substrate Competitive inhibitor Enzyme
Noncompetitive inhibitor (a) Normal binding (b) Competitive inhibition (c) Noncompetitive inhibition
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Enzymes: Function Inhibition Product Inhibition Activators Cofactors
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Product Inhibition Simulation:
- Within each group, designate THREE people as enzymes (foldase1, foldase2, foldase3) and one person as a process. - Team up with two other groups. This gives us three teams. - Each team will have nine enzymes and three people capable of conducting a process.
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First Enzyme: foldase1
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Second Enzyme: foldase2
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Third Enzyme: foldase3
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Completed Airplanes: MUST be IMMEDIATELY returned to foldase1 who must hold them ready for launch. foldase1 must hold completed airplanes on the back of the hand between fingers.
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Completed Airplanes: foldase1 can only hold 8 airplanes (physical maximum) foldase1 may continue its job if able.
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Process: Only ONE process per TEAM can be active at a time unless otherwise specified (take turns). Process uses up completed paper airplanes by taking a completed airplane from a Foldase1 and launching it. The TEAM that manufactures the most airplanes in 4 minutes wins!
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What… …was the substrate? …was the product?
…is the name for foldase2 when it was folding an airplane? …kind of inhibition occurred? …was an active site? …was an allosteric site? …would need to occur so that more airplanes could be made by the foldase enzymes?
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Product Feedback Inhibition
First enzyme in a biosynthesis pathway is(often) an allosteric enzyme Multiple binding sites Binding to effector changes conformation
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Product Feedback Inhibition
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Product Feedback Inhibition
Start of pathway X Inter- mediate Inter- mediate Product Enzyme 1 Enzyme 2 Enzyme 3 Presence of product inhibits enzyme 1
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Enzymes: Function Inhibition Product Inhibition Activators Cofactors
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Activators
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Enzymes: Function Inhibition Product Inhibition Activators Cofactors
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Cofactors Non-protein chemical required for activity of enzymes
Many vitamins are cofactors E.g., Iron, Manganese, Zinc Cofactor functions Assist with chemical reaction Mg2+ and Taq Allow protein to fold E.g., zinc fingers
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Enzymes: Function Inhibition Product Inhibition Activators Cofactors
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Enzymes and Energy
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Enzymes and Energy
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Enzymes in Action: Luciferase
Enzyme that oxidizes a substrate to form light luciferin + O2 → oxyluciferin + light Utilized by multiple organisms Fishes, fungi, copepoda, shrimps, algæ, and more
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Uses of Luciferase Reporter genes Various assays Forensics
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Video Clip http://www.youtube.com/watch?v=UXl8F-eIoiM
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