Exercise 1 - A small polypeptide fragment of the enzyme pyruvate kinase from fug
ID: 51480 • Letter: E
Question
Exercise 1 - A small polypeptide fragment of the enzyme pyruvate kinase from fugu fish was analyzed to determine the primary structure. The following results were obtained:
Trypsin digestion and sequencing led to the following 6 sequences:
o N-I-T-K
o D-A-Y-I-E-K
o V-V-D-V-G-S-K
o C-S-E-E-I-L-W-L-D-Y-K
o I-Y-I-D-D-G-L-I-S-L-Q-V-K
o E-I-G-S-D-Y-L-M-C-E-I-E-N-G-G-T-L
Chymotrypsin digestion and sequencing led to the following 6 sequences:
o D-A-Y
o L-D-Y
o I-E-K-C-S-E-E-I-L-W
o L-M-C-E-I-E-N-G-G-T-L
o K-N-I-T-K-V-V-D-V-G-S-K-I-Y
o I-D-D-G-L-I-S-L-Q-V-K-E-I-G-S-D-Y
3
Exercise 2 - A small polypeptide fragment of histone H4 from mollusk was analyzed to determine the primary structure. The following results were obtained:
Chymotrypsin digestion and sequencing led to the following 5 sequences:
o A-L
o R-I-S-G-L-I-Y
o E-E-T-R-G-V-L-K-V-F
o L-E-N-V-I-R-D-A-V-T-Y
o T-E-H-A-K-C-K-T-V-T-A-M-D-V-V-Y
Endopeptidase V8 (cuts peptide bonds on the carboxyl side of aspartate and glutamate residues) digestion and sequencing led to the following 7 sequences:
o E
o N-V-I-R-D
o V-V-Y-A-L
o A-V-T-Y-T-E
o R-I-S-G-L-I-Y-E
o T-R-G-V-L-K-V-F-L-E
o H-A-K-C-K-T-V-T-A-M-D
Exercise 3 - A small polypeptide fragment of the CFTR protein from chimpanzee was analyzed to determine the primary structure. The following results were obtained:
Trypsin digestion and sequencing led to the following 10 sequences:
o K
o K
o K
o R
o T-K
o Q-S-F-K
o F-S-I-V-Q-K
o Q-T-G-E-F-G-E-K
o N-S-I-L-N-P-I-N-S-I-R
o T-P-L-Q-M-N-G-I-E-E-D
Chymotrypsin digestion and sequencing led to the following 4 sequences:
o K-Q-T-G-E-F
o T-K-K-Q-S-F
o S-I-V-Q-K-T-P-L-Q-M-N-G-I-E-E-D
o G-E-K-R-K-N-S-I-L-N-P-I-N-S-I-R-K-F
Endopeptidase V8 (cuts peptide bonds on the carboxyl side of aspartate and glutamate residues) digestion and sequencing led to the following 5 sequences:
o E
o D
o F-G-E
o T-K-K-Q-S-F-K-Q-T-G-E
o K-R-K-N-S-I-L-N-P-I-N-S-I-R-K-F-S-I-V-Q-K-T-P-L-Q-M-N-G-I-E
Explanation / Answer
electrophoresis of aminoacids based on the polarity towards the electrode .if we are considering the molecular weight and charge seperation with enzymes digestion we get ;
for pyruvate kinase ;
excersise 2; histone 4k ;
excersie 3;
trypsin digestion ; T-P-L-Q-M-N-G-I-E-E-D
chymotrypsin ;K-Q-T-G-E-F
endopeptidase ; T-K-K-Q-S-F-K-Q-T-G-E
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