Exercise 1 - determine the primary structure of the polypeptide One cycle of Edm
ID: 51478 • Letter: E
Question
Exercise 1 - determine the primary structure of the polypeptide
One cycle of Edman degradation yields PTH-Val
Trypsin digestion and sequencing yields the following peptide fragments
o T-N-V-K
o A-A-W-G-K
o V-L-S-P-A-K
o V-G-A-H-A-G-E-Y-G-A-E-A-T-E
Chymotrypsin digestion and sequencing yields the following peptide fragments
o G-A-E-A-T-E
o G-K-V-G-A-H-A-G-E-Y
o V-L-S-P-A-K-T-N-V-K-A-A-W
Exercise 2 - determine the primary structure of the polypeptide
Chymotrypsin digestion and sequencing yields the following peptide fragments
o R-I-P-K-C-R-K-F
o Q-Q-A-Q-H-L-R-A-C-Q-Q-W
o L-G-K-Q-A-N-Q-S-G-G-G-P-S
Trypsin digestion and sequencing yields the following peptide fragments
o K
o R
o C-R
o I-P-K
o A-C-Q-Q-W-L-G-K
o F-Q-Q-A-Q-H-L-R
o Q-A-N-Q-S-G-G-G-P-S
Exercise 3 - determine the primary structure of the protein
Treatment with cyanogen bromide and sequencing yields the following peptide fragments
o D-M
o L-F-M
o R-A-Y-G-N
Chymotrypsin digestion and sequencing yields the following peptide fragments
o G-N
o D-M-L-F
o M-R-A-Y
Explanation / Answer
Exercise 1:
Fragments obtained from Trypsin digestion are as follows:
o T-N-V-K
o A-A-W-G-K
o V-L-S-P-A-K
o V-G-A-H-A-G-E-Y-G-A-E-A-T-E
Fragments obtained from Chymotrypsin digestion are as follows:
o G-A-E-A-T-E
o G-K-V-G-A-H-A-G-E-Y
o V-L-S-P-A-K-T-N-V-K-A-A-W
These two sets of fragments are from the same polypeptide. Therefore, the sequence of the polypeptide can be known from these two sets of fragments.
The Primary structure or the sequence of amino acids of the polypeptide is as follows:
" V-L-S-P-A-K-T-N-V-K-A-A-W-G-K-V-G-A-H-A-G-E-Y-G-A-E-A-T-E "
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