Classroom practice I 1

Classroom practice I: Amino acids and Dissociable group pKa

peptides α-COOH 2
R-COOH (Glu, Asp) 4
AMINO ACIDS, PEPTIDES AND α-NH 3
+
9
R-NH (Arg) +
12
PROTEINS QUESTIONS 2

9. a) Calculate the pI for the following oligopeptide:

Gly-Glu-Asp-Leu-Ala-Arg-Trp-Val.
Answer key provided at the end of the questionnaire b) Given 100 mL of a solution 1 mM of the above
peptide, at pH 4. How many milliliters of NaOH 0,1 M
1. a) Calculate the isoelectric point for glutamic acid. are required to adjust the pH of the solution to 5?
b) Indicate towards which pole this amino acid would Data:
migrate in an electric field at pH 3.22. Dissociable group pKa
c) What about at pH 8.2?. α-COOH 2
Data: pKa = 2.19; pKa = 9.67;. pKa = 4.25
1 2 R R-COOH (Glu, Asp) 4
α-NH 3
+
9
2. Indicate towards which pole the amino acid histidine R-NH (Arg)
2
+
12
would migrate in an electrophoresis performed at:
a) pH 5.0. 10. A protein has the following ionisable groups,
b) pH 7.6. besides the terminal α-amine y α-carboxile: 9 Asp or
Data: pKa = 1.82; pKa = 9.17; pKa = 6.0.
1 2 R
Glu, 3 His, 3 Lys and 3 Arg.
a) Calculate its pI.
3. Calculate the pI for the following amino acids: b) Towards which pole would the protein migrate in a
a) Gly. electrophoresis performed at pH 4?
b) Ser. c) Given 1 mol of the above protein, at pH 1, How
c) Gln. many equivalents of NaOH are required to titrate it?
d) Arg. Data:
Data: Dissociable group pKa
Amino acid pKa 1 pKa 2 pKa R
α-COOH 2
Glycine 2.34 9.60 --
Serine 2.21 9.15 -- α-NH 3
+
9
Glutamine 2.17 9.13 -- R-COOH (Asp, Glu) 4
Arginine 2.17 9.04 12.48 R-NH (His)
+
6
R-NH (Lys)
3
+
11
4. Given a mixture of the following amino acids: Glu, R-NH (Arg)
2
+
12
Ser and Arg,
a) Would it be possible to separate them by 11. Given a solution containing a mixture of the
electrophoresis? following peptides:
b) Which pH would be most suitable?. I) His-Asp-Glu-Glu-Leu
c) Towards which pole (+, -, o) would each amino acid II) Arg-His-Leu-Lys
migrate at that given pH? III) Gly-Ala-Leu-Trp
Data: Amino acid pKa pKa
1 2pKa R
a) Would it be possible to separate them by
Glutamic acid 2.19 9.67 4.25 electrophoresis?
Serine 2.21 9.15 b) Which pH would be most suitable?. Explain the
Arginine 2.17 9.04 12.48 reasoning behind your answer.
Data:
Dissociable group pKa
5. A mixture of Val, Arg, Glu and Ser is subjected to α-COOH 2
electrophoresis at pH 6.0. Predict towards which pole (+, α-NH 3
+
9
-, o) each amino acid would migrate. R-COOH (Asp, Glu) 4
Data: R-NH (His)+
6
Amino acid pKa pKa
1 2 pKa R
R-NH (Lys)3
+
11
Valine 2.32 9.62 -- R-NH (Arg)2
+
12
Arginine 2.17 9.04 12.48 12. Calculate the pI for the following oligopeptide:
Glutamic acid 2.19 9.67 4.25 Gly-Arg-Gly-His-Asp-Asp-Arg-Leu-Asp-Arg-Asp-
Serine 2.21 9,15 -- Asp-Ala.
Data:
6. If a mixture of hemoglobin (pI: 6.8) and Dissociable group pKa
chymotrysinogen (pI: 9.5) is subjected to electrophoresis, α-COOH 2
towards which pole (+, -, o) would every protein migrate R-COOH 4
at pH 6.8 and pH 8?. α-NH 3
+
9
R-NH +
6
7. Which pH would be most suitable to separate a R-NH 2
+
12
mixture of γ-globuline (pI = 6.6), myoglobin (pI = 7) and
hemoglobin (pI = 6.8) by electrophoresis? Explain the 13. a) Calculate the pI for the following oligopeptide:
reasoning behind your answer. Gly-Arg-Gly-Asp-Arg-Arg-Leu-Asp-Arg-Arg-Asp-
Ala.
8. Calculate the pI for the following oligopeptide: Asp- b) How many mL of a 0.2 M solution of HCl are
Arg-Arg-Gly-Arg-Asp-Arg. required to decrease the pH from 13 to 5, in a solution
Data: containing 10 millimoles of the above peptide?.

Degree in Veterinary Medicine. Biochemistry 1

Classroom practice I 2

Data:
Dissociable group pKa
α-COOH 2
R-COOH 4
α-NH 3
+
9
R-NH 2
+
12

14. a) Calculate the pI for the following oligopeptide:

Asp-Gly-Gly-Glu-Lys-Ile-His-Lys-Asp-Asp-Asp-Gln-
Asp
pKa α-COOH = 2; pKa R-COOH = 4; pKa R-His = 6;
pKa α-NH = 9; pKa R-Lys = 11
3
+

b) Which charge would the peptide have at pH 10?

15. a) Calculate the pI for the following oligopeptide:

Glu-Gly-Lys-Gln-Lys-Ile-His-Lys-Asp-Asn-Gly-Glu-
Ile-His-His
pKa α-COOH = 2; pKa R-COOH = 4; pKa R-His = 6;
pKa α-NH = 9; pKa R-NH = 11
3
+
3
+

b) How many mLs of a solution 0.5 M of NaOH is

required to increase the pH from 4 to 8, of a 200 mL of a
2 mM solution of the above peptide?

ANSWER KEY
1. a) pI=3.22; b) It will not move (no charge); c)
Towards the anode (- charged).
2. a) Towards the cathode; b) ) It will not move.
3. a) Gly: 5.97; b) Ser: 5.68; c) Gln: 5.65; d) Arg: 10.76.
4. a) Yes; b) pH 5.68; c) Glu: towards the anode; Ser: it
will not move; Arg: towards the cathode.
5. Val: it will not move; Arg: towards the cathode; Glu:
towards the anode; Ser: slight migration towards the
anode.
6. pH 6.8: Hb will not move and Chtr towards the
cathode; pH 8.0: Hb towards the anode and Chtr towards
the cathode.
7. pH 6.8.
8. pI = 11.52.
9. a) pI = 4; b) 1 ml of 0.1 M NaOH.
10. a) pI = 5.0; b) Towards the cathode; c) 20
equivalents.
11. a) Yes; b) pH 5.5.
12. a) pI = 4.6.
13. a) pI = 11.4; b) 300 mL.
14. a) pI = 4.0; b) Charge –5.
15. a) pI = 7.0; b) 3.6 mL.

Degree in Veterinary Medicine. Biochemistry 2