The document summarizes the derivation of the Michaelis-Menten equation, which describes the relationship between substrate concentration and reaction rate in enzyme-catalyzed reactions. It explains that (1) the steady state assumption states that the rate of enzyme-substrate complex formation equals the rate of complex dissociation, (2) the Michaelis constant km represents the substrate concentration at which the reaction rate is half of its maximum rate Vmax, and (3) km can be used to compare the substrate affinity of different enzymes, with a lower km indicating higher affinity.
The document summarizes the derivation of the Michaelis-Menten equation, which describes the relationship between substrate concentration and reaction rate in enzyme-catalyzed reactions. It explains that (1) the steady state assumption states that the rate of enzyme-substrate complex formation equals the rate of complex dissociation, (2) the Michaelis constant km represents the substrate concentration at which the reaction rate is half of its maximum rate Vmax, and (3) km can be used to compare the substrate affinity of different enzymes, with a lower km indicating higher affinity.
The document summarizes the derivation of the Michaelis-Menten equation, which describes the relationship between substrate concentration and reaction rate in enzyme-catalyzed reactions. It explains that (1) the steady state assumption states that the rate of enzyme-substrate complex formation equals the rate of complex dissociation, (2) the Michaelis constant km represents the substrate concentration at which the reaction rate is half of its maximum rate Vmax, and (3) km can be used to compare the substrate affinity of different enzymes, with a lower km indicating higher affinity.
The document summarizes the derivation of the Michaelis-Menten equation, which describes the relationship between substrate concentration and reaction rate in enzyme-catalyzed reactions. It explains that (1) the steady state assumption states that the rate of enzyme-substrate complex formation equals the rate of complex dissociation, (2) the Michaelis constant km represents the substrate concentration at which the reaction rate is half of its maximum rate Vmax, and (3) km can be used to compare the substrate affinity of different enzymes, with a lower km indicating higher affinity.
𝐸𝐸 + 𝑆𝑆: enzyme reacts with substrate o Through induced fit or lock key model) 𝐸𝐸𝐸𝐸: enzyme-substrate complex formed Place 𝑘𝑘𝑚𝑚 into previous equation 𝐸𝐸 + 𝑃𝑃: will disassociate into enzyme and product [𝐸𝐸𝑇𝑇 ][𝑆𝑆] − [𝐸𝐸𝐸𝐸][𝑆𝑆] = [𝐸𝐸𝐸𝐸] 𝑘𝑘𝑚𝑚 [𝑥𝑥]: concentration of x Addition of [𝐸𝐸𝐸𝐸][𝑆𝑆] (2) Rates [𝐸𝐸𝑇𝑇 ][𝑆𝑆] = [𝐸𝐸𝐸𝐸]𝑘𝑘𝑚𝑚 + [𝐸𝐸𝐸𝐸][𝑆𝑆] 𝑘𝑘1 = rate at which enzyme + substrate is turning into [𝐸𝐸𝑇𝑇 ][𝑆𝑆] = [𝐸𝐸𝐸𝐸](𝑘𝑘𝑚𝑚 + [𝑆𝑆]) enzyme-substrate complex Divide by (𝑘𝑘𝑘𝑘 + [𝑆𝑆]) to solve for [𝐸𝐸𝐸𝐸] 𝑘𝑘−1 = rate at which enzyme-substrate complex [𝐸𝐸𝑇𝑇 ][𝑆𝑆] disassociate back into enzyme + substrate [𝐸𝐸𝐸𝐸] = 𝑘𝑘𝑚𝑚 + [𝑆𝑆] 𝑘𝑘2 = rate at which enzyme-substrate complex disassociates into enzyme + product (2) Initial velocity 𝑽𝑽𝟎𝟎 𝑘𝑘−2 = very small, can be neglected 𝑽𝑽𝟎𝟎 = 𝒌𝒌𝟐𝟐 [𝑬𝑬𝑬𝑬] Solve for [𝐸𝐸𝐸𝐸] 𝑉𝑉0 II) STEADY STATE ASSUMPTION [𝐸𝐸𝐸𝐸] = 𝑘𝑘2 Rate of 𝐸𝐸𝐸𝐸 formation is equal to rate of 𝐸𝐸𝐸𝐸 disassociation [𝐸𝐸𝐸𝐸] equals previous [𝐸𝐸𝐸𝐸] 𝑹𝑹𝑹𝑹𝑹𝑹𝒆𝒆𝑭𝑭 = 𝑹𝑹𝑹𝑹𝑹𝑹𝒆𝒆𝑫𝑫 [𝐸𝐸𝑇𝑇 ][𝑆𝑆] 𝑉𝑉0 = (1) Rate of ES formation 𝑘𝑘2 𝑘𝑘𝑚𝑚 + [𝑆𝑆] Rate of 𝐸𝐸𝐸𝐸 formation Multiply by 𝑘𝑘2 𝑘𝑘1 leads to 𝐸𝐸𝐸𝐸 formation, coming from 𝐸𝐸 + 𝑆𝑆 𝑘𝑘2 [𝐸𝐸𝑇𝑇 ][𝑆𝑆] 𝑉𝑉0 = 𝑘𝑘1 ∗ 𝐸𝐸 𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐 ∗ 𝑆𝑆 𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐 𝑘𝑘𝑚𝑚 + [𝑆𝑆] 𝑅𝑅𝑅𝑅𝑅𝑅𝑒𝑒𝐹𝐹 = 𝑘𝑘1 [𝐸𝐸][𝑆𝑆] (3) Maximal velocity 𝑽𝑽𝒎𝒎𝒎𝒎𝒎𝒎 (2) Rate of ES disassociation Point where all 𝐸𝐸𝐸𝐸 is formed and there is no free 𝐸𝐸 left Rate of 𝐸𝐸𝐸𝐸 disassociation At 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 the [𝐸𝐸𝐸𝐸] = [𝐸𝐸𝑇𝑇 ] because the total enzyme 𝑘𝑘−1 leads to 𝐸𝐸𝐸𝐸 disassociation into 𝐸𝐸 + 𝑆𝑆, coming from 𝐸𝐸𝐸𝐸 concentration is bound to a substrate 𝑘𝑘2 leads to 𝐸𝐸𝐸𝐸 disassociation into 𝐸𝐸 − 𝑃𝑃, coming from 𝐸𝐸𝐸𝐸 𝑘𝑘−1 ∗ 𝐸𝐸𝐸𝐸 𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐 + 𝑘𝑘2 ∗ 𝐸𝐸𝐸𝐸 𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐 𝑅𝑅𝑅𝑅𝑅𝑅𝑒𝑒𝐷𝐷 = 𝑘𝑘−1 [𝐸𝐸𝐸𝐸] + 𝑘𝑘2 [𝐸𝐸𝐸𝐸] (3) Steady State Assumption Rate of 𝐸𝐸𝐸𝐸 formation is equal to rate of 𝐸𝐸𝐸𝐸 disassociation 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 = 𝑘𝑘2 [𝐸𝐸𝑇𝑇 ] Replace 𝑘𝑘2 [𝐸𝐸𝑇𝑇 ] by 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 in equation for 𝑉𝑉0
o This formula is essential for understanding enzyme
kinetics
MICHAELIS MENTEN EQUATION BIOCHEMISTRY: Note #1. 1 of 4
(4) 𝑽𝑽𝟎𝟎 and 𝑽𝑽𝒎𝒎𝒎𝒎𝒎𝒎 together 𝑉𝑉0 = point where 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 is half 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 𝑉𝑉0 = 2 𝑉𝑉0 equals previous 𝑉𝑉0 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 Vmax [𝑆𝑆] = 2 𝑘𝑘𝑚𝑚 + [𝑆𝑆] Solve for 𝑘𝑘𝑚𝑚 , by cross multiplying 2 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 [𝑆𝑆] = 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 (𝑘𝑘𝑚𝑚 + [𝑆𝑆]) Divide by 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 2 [𝑆𝑆] = 𝑘𝑘𝑚𝑚 + [𝑆𝑆] Solve for 𝑘𝑘𝑚𝑚 by subtracting [𝑆𝑆]
𝒌𝒌𝒎𝒎 equals the substrate concentration at half of
maximal rate!
Simplification: 𝑘𝑘𝑚𝑚 tells us the substrate concentration at half 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 𝑘𝑘𝑚𝑚 tells us the affinity of an enzyme for a substrate ↓ 𝒌𝒌𝒎𝒎 = ↑ substrate affinity o Can’t take large amounts of substrate o Example: hexokinase in muscle ↑ 𝒌𝒌𝒎𝒎 = ↓ substrate affinity o Can take larger amounts of substrate o Example: glucokinase in liver Hexokinase and glucokinase are isozymes: enzymes perform same function (add phosphate to 6th carbon of glucose)
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IV) APPENDIX
MICHAELIS MENTEN EQUATION BIOCHEMISTRY: Note #1. 3 of 4
V) REVIEW QUESTIONS
1) What does the steady state assumption say?
a) The rate of ES formation equals the rate of product formation b) The rate of all reactions is equal c) The rate of ES disassociation equals the rate of product formation d) The rate of ES formation equals the rate of ES disassociation 2) When is 𝑽𝑽𝒎𝒎𝒎𝒎𝒎𝒎 reached? a) When all product is formed b) When all enzyme is working on a substrate c) When all enzyme has disassociated from a substrate d) When the steady state assumption is met 3) When does 𝒌𝒌𝒎𝒎 have the same value as the substrate concentration? a) At maximal rate b) At half of maximal rate c) At the end d) When the steady state assumption is met 4) What can a low 𝒌𝒌𝒎𝒎 tell us? a) The affinity of an enzyme to its substrate is low b) The affinity of an enzyme to its substrate is always equal c) The affinity of an enzyme to its substrate is high d) The affinity of an enzyme to its substrate is zero 5) What does a low substrate affinity mean? a) The enzyme can take larger amounts of substrate b) The enzyme can take lesser amounts of substrate c) The enzyme is independent of ATP d) All enzymes can take the same amount of substrate
CHECK YOUR ANSWERS
4 of 4 BIOCHEMISTRY: Note #1. MICHAELIS MENTEN EQUATION
