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    Biochem

    Uploaded by

    Liezyl Dela Cruz
    Michaelis-Menten

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd

    Biochem

    Uploaded by

    Liezyl Dela Cruz
    31 views1 page
    Michaelis-Menten

    Copyright

    © © All Rights Reserved

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    PDF, TXT or read online from Scribd

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    Michaelis-Menten

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    31 views1 page

    Biochem

    Uploaded by

    Liezyl Dela Cruz
    Michaelis-Menten

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 1

    Michaelis-Menten (steady-state) Kinetics

    The Michaelis-Menten model for enzyme kinetics presumes a simple 2-step reaction:
    Step 1: Binding the substrate binds to the enzyme
    Step 2: Catalysis the substrate is converted to product and released
    (Note that enzymes not matching this reaction scheme may still show similar kinetics.)

    The Michaelis-Menten equation shows how the initial rate of


    this reaction, Vo, depends on the substrate concentration, [S]:

    k1
    E+S

    S
    + S

    k2
    ES

    E+P

    k -1

    k-2

    Binding

    Catalysis

    Several simplifying assumptions allow for the derivation of the Michaelis-Menten equation:
    (1)
    (2)
    (3)
    (4)
    (5)

    The binding step ( E + S


    The catalytic step ( ES

    ES ) is fast, allowing the reaction to quickly reach equilibrium ratios of [E], [S], and [ES].
    E + P ) is slower, and thus rate-limiting.
    At early time points, where initial velocity (Vo) is measured, [P] 0.
    ES immediately comes to steady state, so [ES] is constant (throughout the measured portion of the reaction).

    [S] >> [ET], so the fraction of S that binds to E (to form ES) is negligible, and [S] is constant at early time points.
    The enzyme exists in only two forms: free (E), and substrate-bound (ES). Thus, the total enzyme concentration (ET) is the
    sum of the free and substrate-bound concentrations: [ET] = [E] + [ES]

    A derivation of the Michaelis-Menten equation shows how to use the above assumptions to describe the rate of the
    enzyme-catalyzed reaction in terms of measurable quantities:
    From (1), we know the overall rate of the reaction is determined by the rate
    of the catalytic step:

    = ES EP

    From (2), the second term equals zero, so we are left with:

    = ES

    We want to describe Vo in measurable quantities, but [ES] is not easy to


    measure. However [S] is known, from (4). To express [ES] in terms of
    [S], we can start from (3):

    Rate of formation of ES = Rate of breakdown of ES


    ES + EP = ES + ES

    From (2), this simplifies to:

    ES = ES + ES

    We can factor out [ES] and group the rate constants:

    This ratio of rate constants is defined as the Michaelis Constant, Km:


    Substituting in Km for the rate-constant ratio gives:
    Just as [ES] is not easy to measure, [E] is also not easy to measure.
    However, [ET] is known. Rearranging (5) for [E] and substituting, we get:
    We are still trying to get an expression for [ES] in terms of measurable
    quantities. Here we can multiply, rearrange, factor, and divide, to get [ES]
    in terms of [ET], [S], and Km:

    Now we can substitute our expression for [ES] into the rate equation:
    At high [S] (when [S] >>> Km), nearly all enzyme will have substrate
    bound, and [ES] approaches [ET]. This is when Vo approaches Vmax. Since
    Vo = k2[ES],
    (Or, mathematically, when [S] >>> Km, Km is negligible, and the equation
    simplifies to:)
    Substituting Vmax in to the rate equation gives the Michaelis-Menten
    equation:

    ES = ES +
    +
    ES = ES

    ES = ES

    ET ESS = ES
    ET S ESS = ES
    ET S = ES + ESS
    ET S = ES + S
    ET S
    = ES
    + S
    = ES =

    = T
    =
    =

    ET S
    + S

    ET S
    = ET
    S

    S
    S
    + S
    S

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