Purifying a polypeptide, trying to determine the sequence - four consecutive ami

Question

Purifying a polypeptide, trying to determine the sequence - four consecutive amino acid residues permited due to a poor quality sequencer. Biochemical analysis reveals the following:

-Dansyl chloride treatment, followed by acid hydrolysis: dansyl-Pro

-Complete trypsin digestion, follwed by Edman degradation of the separated fragments:

Glu-Lys

Phe-Ile-Val

Pro-Gly-Ala-Arg

Ser-Arg

Provide as much information as you can regarding the primary structure of this polypeptide. Since additional rounds of sequencing are not possible, what additional technique should be used to complete the sequence determination?

Explanation / Answer

trypsin hydrolyses polypeptides on the carboxylic side of arginine and lysine residue.

so the sequence is

Glu-Lys-Phe-Ile- Val-Arg-Gly-Pro-Ser-Arg

other methods to determine additional rounds of sequencing is CNBr fragmentation.

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