Secondary Plot (kinetics)
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Secondary Plot (kinetics)
In enzyme kinetics, a secondary plot uses the intercept or slope from several Lineweaver–Burk plots to find additional kinetic constants. For example, when a set of v by curves from an enzyme with a ping–pong mechanism (varying substrate A, fixed substrate B) are plotted in a Lineweaver–Burk plot, a set of parallel lines will be produced. The following Michaelis–Menten equation relates the initial reaction rate ''v''0 to the substrate concentrations and : \begin \frac &= \frac+\frac+\frac \end The y-intercept of this equation is equal to the following: : \begin \mbox = \frac+\frac \end The y-intercept is determined at several different fixed concentrations of substrate B (and varying substrate A). The y-intercept values are then plotted versus 1/ to determine the Michaelis constant for substrate B, K_M^B, as shown in the Figure to the right.The Horseradish Peroxidase/ o-Phenylenediamine (HRP/OPD) System Exhibits a Two-Step Mechanism. M. K. Tiama and T. M. Hamilton ...
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Enzyme Kinetics
Enzyme kinetics is the study of the rates of enzyme catalysis, enzyme-catalysed chemical reactions. In enzyme kinetics, the reaction rate is measured and the effects of varying the conditions of the reaction are investigated. Studying an enzyme's chemical kinetics, kinetics in this way can reveal the catalytic mechanism of this enzyme, its role in metabolism, how its activity is controlled, and how a drug or a modifier (Enzyme inhibitor, inhibitor or Enzyme activator, activator) might affect the rate. An enzyme (E) is a protein molecule that serves as a biological catalyst to facilitate and accelerate a chemical reaction in the body. It does this through binding of another molecule, its Substrate (biochemistry), substrate (S), which the enzyme acts upon to form the desired product. The substrate binds to the active site of the enzyme to produce an enzyme-substrate complex ES, and is transformed into an enzyme-product complex EP and from there to product P, via a transition state ...
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Enzyme
An enzyme () is a protein that acts as a biological catalyst by accelerating chemical reactions. The molecules upon which enzymes may act are called substrate (chemistry), substrates, and the enzyme converts the substrates into different molecules known as product (chemistry), products. Almost all metabolism, metabolic processes in the cell (biology), cell need enzyme catalysis in order to occur at rates fast enough to sustain life. Metabolic pathways depend upon enzymes to catalyze individual steps. The study of enzymes is called ''enzymology'' and the field of pseudoenzyme, pseudoenzyme analysis recognizes that during evolution, some enzymes have lost the ability to carry out biological catalysis, which is often reflected in their amino acid sequences and unusual 'pseudocatalytic' properties. Enzymes are known to catalyze more than 5,000 biochemical reaction types. Other biocatalysts include Ribozyme, catalytic RNA molecules, also called ribozymes. They are sometimes descr ...
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Lineweaver–Burk Plot
In biochemistry, the Lineweaver–Burk plot (or double reciprocal plot) is a graphical representation of the Michaelis–Menten equation of enzyme kinetics, described by Hans Lineweaver and Dean Burk in 1934. The double reciprocal plot distorts the error structure of the data, and is therefore not the most accurate tool for the determination of enzyme kinetic parameters. While the Lineweaver–Burk plot has historically been used for evaluation of the parameters, together with the alternative linear forms of the Michaelis–Menten equation such as the Hanes–Woolf plot or Eadie–Hofstee plot, all linearized forms of the Michaelis–Menten equation should be avoided to calculate the kinetic parameters. Properly weighted non-linear regression methods are significantly more accurate and have become generally accessible with the universal availability of desktop computers. Definitions The Lineweaver–Burk plot derives from a transformation of the Michaelis–Menten equation ...
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Michaelis–Menten Kinetics
In biochemistry, Michaelis–Menten kinetics, named after Leonor Michaelis and Maud Menten, is the simplest case of enzyme kinetics, applied to enzyme-catalysed reactions involving the transformation of one substrate into one product. It takes the form of a differential equation describing the reaction rate v (rate of formation of product (biology), product P, with concentration p) as a function of a, the concentration of the Enzyme substrate (biology), substrate  A (using the symbols recommended by the IUBMB). Its formula is given by the Michaelis–Menten equation: : v = \frac = \frac V, which is often written as V_\max, represents the limiting rate approached by the system at saturating substrate concentration for a given enzyme concentration. The Michaelis constant K_\mathrm has units of concentration, and for a given reaction is equal to the concentration of substrate at which the reaction rate is half of V. Biochemical reactions involving a single substrate are often ...
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Substrate (biochemistry)
In chemistry, the term substrate is highly context-dependent. Broadly speaking, it can refer either to a chemical species being observed in a chemical reaction, or to a surface on which other chemical reactions or microscopy are performed. In the former sense, a reagent is added to the ''substrate'' to generate a product through a chemical reaction. The term is used in a similar sense in synthetic and organic chemistry, where the substrate is the chemical of interest that is being modified. In biochemistry, an enzyme substrate is the material upon which an enzyme acts. When referring to Le Chatelier's principle, the substrate is the reagent whose concentration is changed. ;Spontaneous reaction : :*Where S is substrate and P is product. ;Catalysed reaction : :*Where S is substrate, P is product and C is catalyst. In the latter sense, it may refer to a surface on which other chemical reactions are performed or play a supporting role in a variety of spectroscopic and mic ...
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Enzyme Inhibitor
An enzyme inhibitor is a molecule that binds to an enzyme and blocks its Enzyme activity, activity. Enzymes are proteins that speed up chemical reactions necessary for life, in which Substrate (biochemistry), substrate molecules are converted into Product (chemistry), products. An enzyme Enzyme catalysis, facilitates a specific chemical reaction by binding the substrate to its active site, a specialized area on the enzyme that accelerates the Rate-determining step, most difficult step of the reaction. An enzyme inhibitor stops ("inhibits") this process, either by binding to the enzyme's active site (thus preventing the substrate itself from binding) or by binding to another site on the enzyme such that the enzyme's catalysis of the reaction is blocked. Enzyme inhibitors may bind Reversible reaction, reversibly or irreversibly. Irreversible inhibitors form a Covalent bond, chemical bond with the enzyme such that the enzyme is inhibited until the chemical bond is broken. By cont ...
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