Question 4 You must cleave the peptide into smaller fragments. Show the exact cl

Question

Question 4 You must cleave the peptide into smaller fragments. Show the exact cleavage sites and the number of resulting peptides after cleavage with the following proteins or chemical reagents. You do each cleavage SEPARATELY, not one after the other. NTQMGRCKPWNTFVHYPLVDVQNVCFKE + Trypsin NTQMGRCKPWNTFVHYPLVDVQNVCFKE + Chymotrypsin NTQMGRCKPWNTFVHYPLVDVQNVCFKE + EndoV8 Question 5 Cleavage of a short, unknown peptide with trypsin yields the following three peptides: Leu-glu; gly-tyr-asn-arg; and gln-ala-phe-val-lys Cleavage of the peptide with chymotrypsin yields the following three peptides: Gln-ala-phe; asn-arg-leu-glu; val-lys-gly-tyr What is the sequence of the peptide?

Explanation / Answer

ANSWER 4-

1) Trypsin cleaves on C- terminal lysine (K) and arginine (R)

So, cleavage will happen at position 6,8 and 27 resulting in 4 smaller peptides.

2) Chemotrypsin cleaves on C- terminal phenylalanine (F), Tryptophan (W) and Tyrosine (Y)

So, cleavage will happen at position 10,13 and 26 resulting in 4 smaller peptides.

3) CNBr cleaves on methionine (M).

So, cleavage will happen at position 4 resulting in 2 smaller peptides.

ANSWER 5-

1) Trypsin cleaves on C- terminal lysine (K) and arginine (R)

Possible sequences are:

2,3,1 i.e., Gly-tyr-asn-arg; gln-ala-phe-val-lys; leu-glu OR

3,2,1 i.e., Gln-ala-phe-val-lys; gly-tyr-asn-arg; leu-glu

2) Chemotrypsin cleaves on C- terminal phenylalanine (F), Tryptophan (W) and Tyrosine (Y)

Possible sequences are:

1,3,2 i.e.,Gln-ala-phe; val-lys-gly-tyr; asn-arg-leu-glu

3,1,2 i.e.,Val-lys-gly-tyr; gln-ala-phe; asn-arg-leu-glu

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