MichaelisMenten kinetics

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An example curve with parametersVmax = 3.4 and Km = 0.4.

In biochemistry, MichaelisMenten kinetics is one of best-known models of enzyme kinetics. It is named after German biochemist Leonor Michaelisand Canadian physician Maud Menten. The model takes the form of an equation describing the rate of enzymatic reactions, by relating reaction rate the concentration of a substrate S. Its formula is given by to ,

. Here, represents the maximum rate achieved by the system, at maximum (saturating) substrate is the substrate concentration at which the reaction rate

concentrations. The Michaelis constant is half of

. Biochemical reactions involving a single substrate are often assumed to follow

MichaelisMenten kinetics, without regard to the model's underlying assumptions.

Contents
[hide]

1 Model 2 Applications 3 Derivation

o o o

3.1 Equilibrium approximation 3.2 Quasi-steady-state approximation 3.3 Assumptions and limitations

4 Determination of constants 5 See also 6 References 7 Further reading

Model[edit]

Change in concentrations over time for enzyme E, substrate S, complex ES and product P

In 1903, French physical chemist Victor Henri found that enzyme reactions were initiated by a bond (more generally, a binding interaction) between the enzyme and the substrate.[1] His work was taken up by German biochemist Leonor Michaelis and Canadian physician Maud Menten, who investigated the kinetics of an enzymatic reaction mechanism, invertase, that catalyzes the hydrolysis of sucrose into glucose and fructose.[2] In 1913, they proposed a mathematical model of the reaction.[3] It involves an enzyme E binding to a substrate S to form a complex ES, which in turn is converted into a product P and the enzyme. This may be represented schematically as

where

, and

denote the rate constants,[4] and the double arrows between S and ES

represent the fact that enzyme-substrate binding is areversible process. Under certain assumptions such as the enzyme concentration being much less than the substrate concentration the rate of product formation is given by

The reaction rate increases with increasing substrate concentration , asymptotically approaching its maximum rate substrate. It also follows that concentration. , attained when all enzyme is bound to , where is the enzyme

, the turnover number, is the maximum number of substrate molecules

converted to product per enzyme molecule per second. The Michaelis constant is the substrate concentration at which the reaction rate is at

half-maximum, and is an inverse measure of the substrate's affinity for the enzymeas a small indicates high affinity, meaning that the rate will approach more

quickly.[5] The value of

is dependent on both the enzyme and the substrate, as well as

conditions such as temperature and pH. The model is used in a variety of biochemical situations other than enzyme-substrate interaction, including antigen-antibody binding, DNA-DNA hybridization, and protein-protein

interaction.[5][6] It can be used to characterise a generic biochemical reaction, in the same way that the Langmuir equation can be used to model generic adsorption of biomolecular species.[6] When an empirical equation of this form is applied to microbial growth, it is sometimes called a Monod equation.

Applications[edit]
Parameter values vary wildly between enzymes:[7]

Enzyme

(M)

(1/s)

(1/M.s)

Chymotrypsin

1.5 102

0.14

9.3

Pepsin

3.0 104

0.50

1.7 103

Tyrosyl-tRNA synthetase

9.0 104

7.6

8.4 103

Ribonuclease

7.9 103

7.9 102

1.0 105

Carbonic anhydrase

2.6 102

4.0 105

1.5 107

Fumarase

5.0 106

8.0 102

1.6 108

The constant

is a measure of how efficiently an enzyme converts a substrate

into product. It has a theoretical upper limit of 108 1010 /M.s; enzymes working close to this, such as fumarase, are termed superefficient.[8] MichaelisMenten kinetics have also been applied to a variety of spheres outside of biochemical reactions,[4] including alveolar clearance of dusts,[9] the richness of species pools,[10] clearance ofblood alcohol,[11] the photosynthesis-irradiance relationship, and bacterial phage infection.[12]

Derivation[edit]
Applying the law of mass action, which states that the rate of a reaction is proportional to the product of the concentrations of the reactants (i.e.[E][S]), gives a system of four nonlinear ordinary differential equations that define the rate of change of reactants with time :[13]

In this mechanism, the enzyme E is a catalyst, which only facilitates the reaction, so that its total concentration, free plus combined, is a constant. This

conservation law can also be observed by adding the first and third equations above.[13][14]

Equilibrium approximation[edit]
In their original analysis, Michaelis and Menten assumed that the substrate is in instantaneous chemical equilibrium with the complex, and thus .[3][14] Combining this relationship with the enzyme

conservation law, the concentration of complex is[14]

where

is the dissociation constant for the enzyme-substrate of the reaction the rate at which P is formed is[14]

complex. Hence the velocity

where

is the maximum reaction velocity.

Quasi-steady-state approximation[edit]
An alternative analysis of the system was undertaken by British botanist G. E. Briggs and British geneticist J. B. S. Haldane in 1925.[15] They assumed that the concentration of the intermediate complex does not change on the time-scale of product formation known as the quasi-steady-state assumption or pseudosteady-state-hypothesis. Mathematically, this assumption means . Combining this relationship

with the enzyme conservation law, the concentration of complex is[14]

where

is known as the Michaelis constant, where

, and

are,

respectively, the constants for substrate unbinding, conversion to product, and binding to the enzyme. Hence the velocity reaction is
[14]

of the

Assumptions and limitations[edit]

The first step in the derivation applies the law of mass action, which is reliant on free diffusion. However, in the environment of a living cell where there is a high concentration of proteins, the cytoplasm often behaves more like a gel than a liquid, limiting molecular movements and altering reaction rates.[16] Whilst the law of mass action can be valid in heterogeneous environments,[17] it is more appropriate to model the cytoplasm as a fractal, in order to capture its limited-mobility kinetics.[18] The resulting reaction rates predicted by the two approaches are similar, with the only difference being that the equilibrium approximation defines the constant as steady-state approximation uses , whilst the quasi. However, each approach

is founded upon a different assumption. The MichaelisMenten equilibrium analysis is valid if the substrate reaches equilibrium on a much faster time-scale than the product is formed or, more precisely, that [14]

By contrast, the BriggsHaldane quasi-steady-state analysis is valid if [13][19]

Thus it holds if the enzyme concentration is much less than the substrate concentration. Even if this is not satisfied, the approximation is valid if is large.

In both the MichaelisMenten and BriggsHaldane analyses, the quality of the approximation improves

as

decreases. However, in model building, Michaelis

Menten kinetics are often invoked without regard to the underlying assumptions.[14] It is also important to remember that, while irreversibility is a necessary simplification in order to yield a tractable analytic solution, in the general case product formation is not in fact irreversible. The enzyme reaction is more correctly described as

In general, the assumption of irreversibility is a good one in situations where one of the below is true: 1. The concentration of substrate(s) is very much larger than the concentration of products:

This is true under standard in vitro assay conditions, and is true for many in vivo biological reactions, particularly where the product is continually removed by a subsequent reaction. 2. The energy released in the reaction is very large, that is

In situations where neither of these two conditions hold (that is, the reaction is low energy and a substantial pool of product(s) exists), the MichaelisMenten equation breaks down, and more complex modelling approaches explicitly taking the forward and reverse reactions into account must be taken to understand the enzyme biology.

Determination of constants[edit]
The typical method for determining the constants and involves

running a series of enzyme assays at varying substrate concentrations measuring the initial reaction rate 'Initial' here is taken to mean that the reaction rate is measured after a relatively short time period, during which it is assumed that the enzyme-substrate complex has formed, but that the substrate concentration held approximately constant, and so the equilibrium or quasi-steadystate approximation remain valid.[19] By plotting reaction rate against concentration, and usingnonlinear regression of the MichaelisMenten equation, the parameters may be obtained.[20] Before computing facilities to perform nonlinear regression became available, graphical methods involving linearisation of the equation were used. A number of these were proposed, including theEadie Hofstee diagram, HanesWoolf plot and LineweaverBurk plot; of these, the HanesWoolf plot is the most accurate.[20] However, while useful for visualization, all three methods distort the error structure of the data and are inferior to nonlinear regression.[21] Nonetheless, their use can still be found in modern literature.[22] In 1997 Santiago Schnell and Claudio Mendoza derived a closed form solution for the time course kinetics analysis of the MichaelisMenten kinetics.[23] The solution, known as the Schnell-Mendoza equation, has the form: , and .

where W[] is the Lambert-W function and where F(t) is

The Schnell-Mendoza equation has been used to estimate and

[24][25]

from

time course data.

The equation below, obtained by Berberan-Santos in 2010 (MATCH Commun. Math. Comput. Chem. 63, 283), encompasses the SchnellMendoza equation, and is still valid when the initial substrate concentration is close to that of enzyme,

where W[] is again the Lambert-W function.