What is the charge on lysine at pH 7.0?
The pKa values are as follows:
a) carboxyl at C terminus – 2.0
b) amino at N terminus – 9.0
c) Lys side chain – 10.0
pH = pKa + log (A-/HA) Charge = “# charge”/(“# charge” + “# neutral”)
(the sign of this charge depends on the ionizable group)
for a) 7 = 2 + log (COO-/COOH)
5 = log (COO-/COOH)
105 = (COO-/COOH)
(COO-/COOH) = 100,000/1 Charge = 100,000/(100,000 + 1) = 1.0 ŕ -1.0
for b) 7 = 9 + log (NH2/NH3+)
-2 = log (NH2/NH3+)
10-2 = (NH2/NH3+)
(NH2/NH3+) = 1/102 = 1/100 Charge = 100/(100 + 1) = 1.0 ŕ +1.0
for c) 7 = 10 + log (NH2/NH3+)
-3 = log (NH2/NH3+)
10-3 = (NH2/NH3+)
(NH2/NH3+) = 1/103 = 1/1,000
Charge = 1,000/(1,000 + 1) = 1.0 ŕ +1.0
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TOTAL CHARGE = + 1.0
What is the charge on lysine at pH 10.0?
The pKa values are as follows:
a) carboxyl at C terminus – 2.0
b) amino at N terminus – 9.0
c) Lys side chain – 10.0
pH = pKa + log (A-/HA) Charge = “# charge”/(“# charge” + “# neutral”)
(the sign of this charge depends on the ionizable group)
for a) 10 = 2 + log (COO-/COOH)
8 = log (COO-/COOH)
108 = (COO-/COOH)
(COO-/COOH) = 100,000,000/1
Charge = 100,000,000/(100,000,000 + 1) = 1.0 ŕ -1.0
for b) 10 = 9 + log (NH2/NH3+)
1 = log (NH2/NH3+)
101 = (NH2/NH3+)
(NH2/NH3+) = 10/1 Charge = 1/(10 + 1) = 1/11 = 0.1 ŕ +0.1
for c) 10 = 10 + log (NH2/NH3+)
0 = log (NH2/NH3+)
100 = (NH2/NH3+)
(NH2/NH3+) = 1 = 1/1 Charge = 1/(1+ 1) = 0.5 ŕ +0.5
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TOTAL CHARGE = -0.4
Consider the following peptide:
Glu-Ala-Arg-Ser-Tyr
What is the net charge (to 0.1 unit) on the peptide at pH 3.0?
Assume the pKa values are as follows:
a) carboxyl at C terminus – 3.0
b) Glu side chain – 4.0
c) amino at N terminus – 8.0
d) Tyr side chain – 10.0
e) Arg side chain – 12.0
for a) 3 = 3 + log (COO-/COOH)
0 = log (COO-/COOH)
100 = (COO-/COOH)
(COO-/COOH) = 1 = 1/1 Charge = 1/(1+1) = 0.5 ŕ -0.5
for b) 3 = 4 + log (COO-/COOH)
-1 = log (COO-/COOH)
10-1 = (COO-/COOH)
(COO-/COOH) = 0.1 = 1/10 Charge = 1/(1+10) = 0.1 ŕ -0.1
for c) 3 = 8 + log (NH2/NH3+)
-5 = log (NH2/NH3+)
10-5 = (NH2/NH3+)
(NH2/NH3+) = 1/105 = 1/100,000 Charge = 100,000/(100,000 + 1) = 1.0 ŕ +1.0
for d) 3 = 10 + log (-O-/-OH)
-7 = log (-O-/-OH)
10-7 = (-O-/-OH)
(-O-/-OH) = 1/107 = 1/10,000,000 Charge = 1/(1 + 10,000,000 = 0.0 ŕ 0.0
For e) 3 = 12 + log (=NH/=NH2+)
-9 = log (=NH/=NH2+)
10-9 = (=NH/=NH2+)
(=NH/=NH2+) = 1/109 = 1/1,000,000,000
Charge = 1,000,000,000/(1,000,000,000 + 1) = 1.0 ŕ +1.0
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TOTAL CHARGE = + 1.4
At pH 7, charges for a) through e) are:
-1.0, -1,0, +0.9, 0, +1.0 to give a total charge of -0.1 (close to the pI)
At pH 10, charges for 1) through e) are:
-1.0, -1.0, 0, -0.5, +1.0 to give a total charge of -1.5