In: Biology
Question) How do we calculate the isoelectric point (PI) of peptides?
so here the given peptide contains 3 aminoacids- (lysine-glutamate-aspartate)
so first we have to estimate the pH at which the net charge on the protein is zero. for that we have to consider the pKa values of alpha- carboxyl group (here it is 1.99), alpha-amino group (given, 9.06) as well as the pKa values of the side chains of the aminoacids. (for that refer the image)
so lets assume the pH as 7, and find out which side chains are protonated (+ve value) and deprotonated (-ve value)
protonated if pKa value is greater than pH value
deprotonated if pKa value is lesser than pH value
aminogroup of lysine (Lys) |
side chain of lysine (Lys) | side chain of glutamate (Glu) | side chain of aspartate (Asp) | carboxyl group of aspartate (Asp) |
pKa 9.06(given) | pKa 10.5 | pKa 4.2 | pKa3.9 | pKa1.99 (given) |
+ve | +ve | -ve | -ve | -ve |
so, +1+1-1-1-1 = -1 (so the net charge on the protein is not zero)
lets assume the pH value as 4.2 . So the side chain of glutamate will be neutralised.
aminogroup of lysine (Lys) |
side chain of lysine (Lys) | side chain of glutamate (Glu) | side chain of aspartate (Asp) | carboxyl group of aspartate (Asp) |
pKa 9.06 (given) | pKa 10.5 | pKa 4.2 | pKa 3.9 | pKa1.99 (given) |
+ve | +ve | 0 | -ve |
-ve |
so, +1+1+0-1-1= 0 (so at pH 4.2 the net charge on the protein is zero)
next step is to calculate the average of the two pKa values directly above and directly below the estimated pH.
that is 9.06 and 3.9 which are directly above and directly below the estimated pH 4.2
so the average or the isoelectric point of the given peptide Lys-Glu-Asp is = (9.06+3.9)/2
= 6.48
Option d (6.48) is the right answer