#CALCULATE PI ISOELECTRIC POINT HOW TO#

To balance the negative charge in the side chain, carboxylate ion has to be protonated. This is because at pH 7 the acidic amino acid’s side chain carboxylic acid group exists as a carboxylate ion, so it has a net negative charge. Amino acids having acidic side chains have isoelectric points less than 7.

The isoelectric point of neutral amino acids is near pH 7.

The isoelectric point of each amino acid is different due to the presence of a unique side chain group, usually represented with the letter R.

Sidechain groups present in an amino acid determine the isoelectric point of that amino acid.

The isoelectric point of amino acids differs based on the following factors: PI=8.95+10.532=9.74 Isoelectric points of acidic and basic amino acids To calculate the isoelectric point of Lysine, the pKa of the similarly ionizable groups will be considered which are the ammonium group (pKa = 8.95) and the basic side chain (pKa = 10.53). Lysine has a basic group in its side chain.

Calculate the isoelectric point of Lysine.

Since Alanine has a neutral side chain (-CH3), the isoelectric point will be calculated using the pKa values of the carboxylic acid group and the ammonium group:

Calculate the isoelectric point of Alanine.

The amino acids exist as zwitterions in an aqueous solution that has a neutral pH.Ī diagram representing the effect of pH on the net charge of a neutral amino acid. This means that at the isoelectric point, the positively charged groups in the solution are balanced by the negatively charged groups. The isoelectric point, pI, is defined as the pH of an aqueous solution of an amino acid at which the amino acids in the solution carry zero net charge. This will be explained further in the examples section. For amino acids having a charged or ionizable side chain, the pI value is calculated by taking a mean of the pKas’ of similarly ionizable groups. The isoelectric point formula for an amino acid having a neutral side chain will be as follows:įor amino acids having a neutral side chain, the pI value is calculated by taking a mean of the pKa of the carboxylate and the pKa of the aminonium group. The positively charged ammonium balances the negative charge of the carboxylate. The carboxyl group loses its hydrogen that is taken up by the amino group to form ammonium. At a neutral pH, an acid-base reaction occurs between the carboxyl group and the amino group of the amino acid. Amino acids interact intramolecularly to form zwitterions. At this pH, the aminoacids show minimum solubility. In an aqueous solution having a pH equal to the isoelectric point, most amino acids are present in the solution as zwitterions. The two opposite charges cancel out giving the zwitterion a net charge of zero. The zwitterion is the electrically neutral form of amino acid, that carries a negatively charged carboxylate group (CO2 –) and a positively charged amino group (NH3+ ). An aminoacid carrying no net charge is called a zwitterion. The isoelectric point is the pH of a solution at which an amino acid or peptide is neutral and carries zero net electric charge. This article will explain what is the isoelectric point, the formula used to calculate isoelectric point, and examples to demonstrate how the formula may be applied. For instance, in isoelectric focusing, a pH gradient is used to separate proteins based on their isoelectric points. The isoelectric point of proteins plays a significant role in the separation and purification methods used to isolate proteins from a solution. The pI value helps you determine the solubility of a molecule at a given pH. The isoelectric point is the pH at which the net electrical charge on a molecule is zero.

#CALCULATE PI ISOELECTRIC POINT HOW TO#

An understanding of how to calculate isoelectric point is essential when you are working with proteins.

Calculate pi isoelectric point