Protein sequencing Edman degradation

The general idea of peptide sequencing by Edman degradation is to cleave one amino acid at a time from an end of the peptide chain. That terminal amino acid is then separated and identified, and the cleavage reactions are repeated on the chain-shortened peptide until the entire peptide sequence is known. Automated protein sequencers are available that allow as many as 50 repetitive sequencing cycles to be carried out before a buildup of unwanted by products interferes with the results. So efficient are these instruments that sequence information can be obtained from as little as 1 to 5 picomoles of sample—less than 0.1 /xg. 

Recombinant DNA Although numerous proteins have been sequenced by Edman degradation using technology the above strategy, the determination of the sequences of large proteins by this... 

Hundreds of proteins have been sequenced by Edman degradation of peptides derived from specific cleavages. Nevertheless, heroic effort is required to elucidate the sequence of large proteins, those with more than 1000 residues. 

Miyashita M, Presley JM, Buchholz BA, Lam KS, Lee YM, Vogel JS, Hammock BD. Attomole level protein sequencing by Edman degradation coupled with accelerator mass spectrometry. Proc Natl Acad Sci U.S.A. 2001 98(8) 4403—4408. 

Figure 16 Schematic representation of an approach to identify proteins separated by ID or 2D gel electrophoresis including MALDI-TOF and ESI-MS/MS followed by database search. Reversed-phase micro-HPLC and automated protein sequencing by Edman degradation are complementary techniques that may prove useful depending on the number of samples to be analyzed and the estimated amount.

N-terminal sequencing by Edman degradation is frequently used to determine the N-terminal amino acid sequence of a protein. The peptide bonds are sequentially hydrolyzed from the N-terminus, and the released amino acid is... 

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