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Be sure to answer all parts. An octapeptide contains the following amino acids: Ala, Pro, Gly, Asp, Trp, Lys, Leu, Met. Carbo

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Carboxypeptidase is an enzyme that hydrolyzes a peptide bond at the carboxy-terminal (C-terminal) at the end of a peptide.

  • Accordingly, ‘Asp’ is the ‘C-terminal’ of the octapeptide.
  • So, 1-2-3-4-5-6-7-Asp

Edman degradation method is used to find the sequence of amino acids in a peptide. This method cleaves the peptide from N-terminal without disturbing the peptide bonds between other amino acid.

  • Since ‘Gly’ comes out of octapeptide, ‘Gly’ is the ‘N’ terminal of octapeptide.             i.e. Gly-2-3-4-5-6-7-Asp
  • Since ‘Gly’ comes out of tetrapeptide ‘A’, ‘Gly’ is the ‘N’ terminal of ‘A’                       i.e. A = Gly-2-3-4   
  • Since ‘Met’ comes out of tetrapeptide ‘B’, ‘Met’ is the ‘N’ terminal of ‘B’                    i.e. B = Met-6-7-Asp
  • Since ‘Gly’ comes out of dipeptide ‘C’, ‘Gly’ is the ‘N’ terminal of ‘C’                            i.e. C = Gly-2
  • Since ‘Leu’ comes out of dipeptide ‘D’, ‘Leu’ is the ‘N’ terminal of ‘D’                            i.e. D = Leu-4

Chymotrypsin is an enzyme that cleaves on the C-terminal phenylalanine (Phe), tryptophan (Trp), and tyrosine (Tyr) on peptide chains. It shows specificity for aromatic amino acids because of its hydrophobic pocket.

  • Accordingly, octapeptide breaks at ‘Trp’ forming two tetrapeptides ‘A’ & ‘B’.
  • So, Gly-2-Leu-Trp-Met-6-7-Asp ---> Gly-2-Leu-Trp (A) + Met-6-7-Asp (B)

Trypsin is an enzyme that cleaves peptide chains mainly at the carboxyl side of the amino acids lysine or arginine.

  • So, position ‘2’ must be ‘Lysine-Lys’
  • Gly-Lys-Leu-Trp (A) + trypsin --> Gly-Lys (C) + Leu-Trp (D)

Partial hydrolysis of B:

Since Met-Pro is one of the products, position ‘6’ must be ‘Pro’.

Met-Pro-7-Asp (B) ---> Met-Pro + other product

Since ‘Ala’ is the only amino acid left, that is bound to take the position ‘7’.

So the octapeptide is: Gly-Lys-Leu-Trp-Met-Pro-Ala-Asp

A = Gly-Lys-Leu-Trp

B = Met-Pro-Ala-Asp

C= Gly-Lys

D= Leu-Trp

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