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The pKa values and the isoelectronic point, pI, are given below for the 20 amino acids.
pKa1= -carboxyl group, pKa2 =-ammonium ion, and pKa3 = side chain
group.
Amino acid
Glycine
Alanine
Valine
Leucine
Isoleucine
Methionine
Proline
Phenylalanine
Tryptophan
Asparagine
Glutamine
Serine
Threonine
Tyrosine
Cysteine
Aspartic acid
Glutamic acid
Lysine
Arginine
Histidine
pKa1
2.34
2.34
2.32
2.36
2.36
2.28
1.99
1.83
2.83
2.02
2.17
2.21
2.09
2.20
1.96
1.88
2.19
2.18
2.17
1.82
pKa2
9.60
9.69
9.62
9.60
9.60
9.21
10.60
9.13
9.39
8.80
9.13
9.15
9.10
9.11
8.18
9.60
9.67
8.95
9.04
9.17
pKa3
------------------------------3.65
4.25
10.53
12.48
6.00
pI
5.97
6.00
5.96
5.98
6.02
5.74
6.30
5.48
5.89
5.41
5.65
5.68
5.60
5.66
5.07
2.77
3.22
9.74
10.76
7.59
Amino Acid
-NH3+
pI
Glycine, Gly
-H
9.78
2.35
5.97
Alanine, Ala
-CH3
9.87
2.35
6.02
Valine, Val
-CH(CH3)2
9.74
2.29
5.97
Leucine, Leu
CH2CH(CH3)2
9.74
2.33
5.98
Isoleucine, Ile
CH(CH3)CH2CH3 9.76
2.32
6.02
Phenylalanine, Phe
9.31
2.20
5.48
Tryptophan, Trp
9.41
2.46
5.88
Tyrosine, Tyr
9.21
2.20
10.46
5.65
Histidine, His
9.33
1.80
6.04*
7.58
Serine, Ser
CH2OH
9.21
2.19
5.68
Threonine, Thr
CH(CH3)-OH
9.10
2.09
6.53
Methionine, Met
CH2CH2SCH3
9.28
2.13
5.75
Cysteine, Cys
CH2SH
10.70
1.92
8.37
5.14
CH2CO2H
9.90
1.99
3.90
2.87
CH2CH2CO2H
9.47
2.10
4.07
3.22
Asparagine, Asn
CH2CONH2
8.72
2.14
5.41
Glutamine, Gln
CH2CH2CONH2
9.13
2.17
5.65
Lysine, Lys
(CH2)4NH2
9.06
2.16
10.54*
9.74
8.99
1.82
12.48*
10.76
Arginine, Arg
Proline, Pro
10.64
1.95
6.10
Isoelectronic point, pI
The isoelectronic point or isoionic point is the pH at which the amino acid
does not migrate in an electric field.
This means it is the pH at which the amino acid is neutral, i.e. the zwitterion
form is dominant.
A table of pKa and pI values can be found on the next page.
The pI is given by the average of the pKas that involve the zwitterion, i.e. that
give the boundaries to its existence.
There are 3 cases to consider....
neutral side chains
These amino acids are characterised by two pKas : pKa1 and pKa2 for the carboxylic
acid and the amine respectively.
The isoelectronic point will be halfway between, or the average of, these two
pKas, i.e. pI = 1/2 (pKa1 + pKa2). This is most readily appreciated when you realise
that at very acidic pH (below pKa1) the amino acid will have an overall +ve charge
and at very basic pH (above pKa2 ) the amino acid will have an overall -ve charge.
For the simplest amino acid, glycine, pKa1= 2.34 and pKa2 = 9.6, pI = 5.97.
The other two cases introduce other ionisable groups in the side chain "R" described
by a third acid dissociation constant, pKa3
acidic side chains
The pI will be at a lower pH because the acidic side chain introduces an "extra"
negative charge. So the neutral form exists under more acidic conditions when the
extra -ve has been neutralised. For example, for aspartic acid shown below, the
neutral form is dominant between pH 1.88 and 3.65, pI is halfway between these two
values, i.e. pI = 1/2 (pKa1 + pKa3), so pI = 2.77.
1. Introduction
Chemical properties of molecules depend largely on whether they are ionized
or not. Most organic molecules are capable of gaining and/or losing a proton
in aqueous solutions. Proton transfer most frequently occurs between water
and any ionizable atom of the organic molecule. The molecule's response to
protonation or deprotonation depends significantly on the site that was
disturbed by proton transfer. Partial charge distribution in the molecule also
varies with protonation of the acid/base active sites. Since the partial charge
distribution is very sensitive to the protonation-deprotonation process (both
near and far from the disturbed site), it can be used to determine the pK of
a molecule. Our pK prediction program is based on the calculation of partial
charge of atoms in the molecule.
a
3. Definition of pKa
When the pH of the solution is equal with pK , the concentrations of dissociated and
undissociated species are equal.
a
5. Multiprotic Molecules
When a molecule contains more than one ionizable atom, it is called a multiprotic
compound. For these types of molecules we need to distinguish between micro and
macro acidic dissociation constants. The micro acidic dissociation constant is
obtained from the equilibrium concentration of the conjugated acid-base pairs. The
macro acidic dissociation constant is obtained from the global mass and charge
conservation law. When a molecule has N ionizable sites, the total number of micro
species in the solution is 2 .
N
charge
AH H H
AH H , AH H , AH H
1
AH , AH , AH
1
-1
-2
-3
The AH molecule has three macro acidic constants since it is triprotic acid. Macro
acidic constants K , K , and K are obtained from the concentration of the
microspecies:
3
And the three macro pK values of the AH molecule that would be obtained with
routine laboratory measurements are as follows:
a
pK =-log (K )
pK =-log (K )
pK =-log (K )
a,1
10
a,2
10
a,3
10
7. Examples
Example 1.
The ionization steps of p-amino benzoic acid are outlined below. Calculated micro
ionization constants k , k , k and k are indicated on the arrows:
1
Below are the calculated and the experimental pK of p-amino benzoic acid
a
Example 2.
Imides and amides can have either acidic or basic character. The extent of
amide/imide ionization at a given pH is determined by two acid dissociation
constants:
pK assigned to the deprotonation step
a,1
RNH
RN + H
-
RNH
+
2
RNH + H
a,2
a,2
a,1
a,2
Chemists often want to know the ionization state of organic compounds at pH 7.4.
In general, the macro pK s of amide/imide compounds are calculated and their
acidic or basic character determined with the above formulas at pH 7.4.
a
2.
phtalimide
8.22
8.30
2-pyridone
11.40
11.70
Example 3
20
The molecule depicted below contains a very weak basic atom. First, macro
pK is calculated with default limits predefined between 10 and 10 . Then
macro pK is calculated with altered limits defined between 10 and 10 .
Changing of the default settings of macro pK calculation can be done in the
-10
20
-50
20
Tools
Options
pK menu of MarvinSketch.
Calculated and observed acidity constants are summarized in Table 3.
Measured pK s taken from Ref.3.
a
First calc.
Second calc.
Observed
pK
a,1
4.24
4.24
4.51
pK
5.75
5.75
6.01
pK
-35.81
a,2
a,3