Dissociation constant
In chemistry, biochemistry, and pharmacology, a dissociation constant (K_{d} ) is a specific type of equilibrium constant that measures the propensity of a larger object to separate (dissociate) reversibly into smaller components, as when a complex falls apart into its component molecules, or when a salt splits up into its component ions. The dissociation constant is the inverse of the association constant. In the special case of salts, the dissociation constant can also be called an ionization constant.
For a general reaction:
 \mathrm{A}_{x}\mathrm{B}_{y} \rightleftharpoons x\mathrm{A} + y\mathrm{B}
in which a complex \mathrm{A}_{x}\mathrm{B}_{y} breaks down into x A subunits and y B subunits, the dissociation constant is defined
 K_{d} = \frac
where [A], [B], and [A_{x}B_{y}] are the concentrations of A, B, and the complex A_{x}B_{y}, respectively.
One reason for the popularity of the dissociation constant in biochemistry and pharmacology is that in the frequently encountered case where x=y=1, K_{d} has a simple physical interpretation: when [A]=K_d, [B]=[AB] or equivalently [AB]/([B]+[AB])=1/2. That is, K_{d}, which has the dimensions of concentration, equals the concentration of free A at which half of the total molecules of B are associated with A. This simple interpretation does not apply for higher values of x or y. It also presumes the absence of competing reactions, though the derivation can be extended to explicitly allow for and describe competitive binding. It is useful as a quick description of the binding of a substance, in the same way that EC50 and IC50 describe the biological activities of substances.
Contents

Concentration of bound molecules 1
 Molecules with one binding site 1.1
 Macromolecules with identical independent binding sites 1.2

Proteinligand binding 2
 Antibodies 2.1
 Acid–base reactions 3
 Dissociation constant of water 4
 See also 5
 References 6
Concentration of bound molecules
Molecules with one binding site
Experimentally, the concentration of the molecule complex [AB] is obtained indirectly from the measurement of the concentration of a free molecules, either [A] or [B]. ^{[1]} In principle, the total amounts of molecule [A]_{0} and [B]_{0} added to the reaction are known. They separate into free and bound components according to the mass conservation principle:
 [A]_0 = [A] + [AB]
 [B]_0 = [B] + [AB]
To track the concentration of the complex [AB], one substitutes the concentration of the free molecules ([A] or [B]), of the respective conservation equations, by the definition of the dissociation constant,
 [A]_0 = K_d \frac + [AB]
This yields the concentration of the complex related to the concentration of either one of the free molecules
 [AB] = \frac{K_d + [B]} = \frac{K_d + [A]}
Macromolecules with identical independent binding sites
Many biological proteins and enzymes can possess more than one binding site. ^{[1]} Usually, when a ligand L binds with a macromolecule M, it can influence binding kinetics of other ligands L binding to the macromolecule. A simplified mechanism can be formulated if the affinity of all binding sites can be considered independent of the number of ligands bound to the macromolecule. This is valid for macromolecules composed of more than one, mostly identical, subunits. It can be then assumed that each of these n subunits are identical, symmetric and that they possess only one single binding site. Then, the concentration of bound ligands [L]_{bound} becomes
 [L]_{bound} = \frac{n[M]_0 [L]}{K_d + [L]}
In this case, [L]_{bound} \neq [LM], but comprises all partially saturated forms of the macromolecule:
 [L]_{bound} = [LM] + 2[L_2 M] + 3[L_3 M] + ... + n[L_n M]
where the saturation occurs stepwise
 [L] + [M] \rightleftharpoons [LM] \qquad K'_1 = \frac \qquad [LM] = \frac{K'_1}
 [L] + [LM] \rightleftharpoons [L_2 M] \qquad K'_2 = \frac \qquad [L_2 M] = \frac{K'_1 K'_2}
 [L] + [L_2 M] \rightleftharpoons [L_3 M] \qquad K'_3 = \frac \qquad [L_3 M] = \frac{K'_1 K'_2 K'_3}
 \vdots
 [L] + [L_{n1} M] \rightleftharpoons [L_n M] \qquad K'_n = \frac \qquad [L_n M] = \frac{K'_1 K'_2 K'_3 \cdots K'_n}
For the derivation of the general binding equation a saturation function r is defined as the quotient from the portion of bound ligand to the total amount of the macromolecule:
 r = \frac = \frac{\sum_{i=1}^n \left( \frac{i [L]^i}{\prod_{j=1}^i K_j'} \right) }{1 + \sum_{i=1}^n \left( \frac{K_d} \right)^i }{1 + \sum_{i=1}^n \left( \prod_{j=1}^i \frac{nj+1}{j} \right) \left( \frac{K_d} \right)^i} = \frac{\sum_{i=1}^n i \binom{n}{i} \left( \frac{K_d} \right)^i }{1 + \sum_{i=1}^n \binom{n}{i} \left( \frac{K_d} \right)^i}
where \binom{n}{i} = \frac{n!}{(ni)!i!} is the binomial coefficient. Then, the first equation is proved by applying the binomial rule
 r = \frac{n \left( \frac{K_d} \right) \left(1 + \frac{K_d} \right)^{n1} }{\left(1 + \frac{K_d} \right)^n} = \frac{n \left( \frac{K_d} \right) }{\left(1 + \frac{K_d} \right)} = \frac{n [L]}{K_d + [L]} = \frac{\left[ \mathrm{LP} \right]}
where [\mathrm{P}], [\mathrm{L}] and [\mathrm{LP}] represent molar concentrations of the protein, ligand and complex, respectively.
The dissociation constant has molar units (M), which correspond to the concentration of ligand [\mathrm{L}] at which the binding site on a particular protein is half occupied, i.e. the concentration of ligand, at which the concentration of protein with ligand bound [\mathrm{C}], equals the concentration of protein with no ligand bound [\mathrm{P}]. The smaller the dissociation constant, the more tightly bound the ligand is, or the higher the affinity between ligand and protein. For example, a ligand with a nanomolar (nM) dissociation constant binds more tightly to a particular protein than a ligand with a micromolar (μM) dissociation constant.
Subpicomolar dissociation constants as a result of noncovalent binding interactions between two molecules are rare. Nevertheless, there are some important exceptions. Biotin and avidin bind with a dissociation constant of roughly 10^{15} M = 1 fM = 0.000001 nM.^{[4]} Ribonuclease inhibitor proteins may also bind to ribonuclease with a similar 10^{15} M affinity.^{[5]} The dissociation constant for a particular ligandprotein interaction can change significantly with solution conditions (e.g. temperature, pH and salt concentration). The effect of different solution conditions is to effectively modify the strength of any intermolecular interactions holding a particular ligandprotein complex together.
Drugs can produce harmful side effects through interactions with proteins for which they were not meant to or designed to interact. Therefore much pharmaceutical research is aimed at designing drugs that bind to only their target proteins (Negative Design) with high affinity (typically 0.110 nM) or at improving the affinity between a particular drug and its invivo protein target (Positive Design).
Antibodies
In the specific case of antibodies (Ab) binding to antigen (Ag), usually the affinity constant is used. It is the inverted dissociation constant.
 \text{Ab} + \text{Ag} \rightleftharpoons \text{AbAg}
 K_{a} = \frac{\left[ \text{AbAg} \right]}{\left[ \text{Ab} \right] \left[ \text{Ag} \right]} = \frac{1}{K_{d}}
This chemical equilibrium is also the ratio of the onrate (k_{forward}) and offrate (k_{back}) constants. Two antibodies can have the same affinity, but one may have both a high on and offrate constant, while the other may have both a low on and offrate constant.
 K_{a} = \frac{k_\text{forward}}{k_\text{back}} = \frac{\mbox{onrate}}{\mbox{offrate}}
Acid–base reactions
Acids and bases 


Acid types 
Base types 

For the deprotonation of acids, K is known as K_{a}, the acid dissociation constant. Stronger acids, for example sulfuric or phosphoric acid, have larger dissociation constants; weaker acids, like acetic acid, have smaller dissociation constants.
(The symbol K_{a}, used for the acid dissociation constant, can lead to confusion with the association constant and it may be necessary to see the reaction or the equilibrium expression to know which is meant.)
Acid dissociation constants are sometimes expressed by pK_{a}, which is defined as:
 \mathrm{p}K_{a} = \log_{10}{K_{a}}
This \mathrm{p}K notation is seen in other contexts as well; it is mainly used for covalent dissociations (i.e., reactions in which chemical bonds are made or broken) since such dissociation constants can vary greatly.
A molecule can have several acid dissociation constants. In this regard, that is depending on the number of the protons they can give up, we define monoprotic, diprotic and triprotic acids. The first (e.g. acetic acid or ammonium) have only one dissociable group, the second (carbonic acid, bicarbonate, glycine) have two dissociable groups and the third (e.g. phosphoric acid) have three dissociable groups. In the case of multiple pK values they are designated by indices: pK_{1}, pK_{2}, pK_{3} and so on. For amino acids, the pK_{1} constant refers to its carboxyl (COOH) group, pK_{2} refers to its amino (NH_{3}) group and the pK_{3} is the pK value of its side chain.
H_3 B \rightleftharpoons\ H ^ + + H_2 B ^  \qquad K_1 = \qquad pK_1 =  \log K_1
H_2 B ^  \rightleftharpoons\ H ^ + + H B ^ {2} \qquad K_2 = \over [H_2 B^ ]} \qquad pK_2 =  \log K_2
H B ^{2} \rightleftharpoons\ H ^ + + B ^{3} \qquad K_3 = \over [H B ^ {2}]} \qquad pK_3 =  \log K_3
Dissociation constant of water
The dissociation constant of water is denoted K_{w}:
K_w = [\mbox{H}^+] [\mbox{OH}^]
The concentration of water \left[ \mbox{H}_2\mbox{O} \right] is omitted by convention, which means that the value of K_{w} differs from the value of K_{eq} that would be computed using that concentration.
The value of K_{w} varies with temperature, as shown in the table below. This variation must be taken into account when making precise measurements of quantities such as pH.
Water temperature  K_{w} / 10^{−14}  pK_{w}^{[6]} 

0 °C  0.112  14.95 
25 °C  1.023  13.99 
50 °C  5.495  13.26 
75 °C  19.95  12.70 
100 °C  56.23  12.25 
See also
References
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