The Edman Degradation Reaction
The peptide to be sequenced is adsorbed onto a solid surface - one common substrate is glass fibre coated with polybrene, a cationic polymer. The Edman reagent, phenylisothiocyanate (PITC), is added to the adsorbed peptide, together with a mildly basic buffer solution of 12% trimethylamine. This reacts with the amine group of the N-terminal amino acid.
The terminal amino acid can then be selectively detached by the addition of anhydrous acid. The derivative then isomerises to give a substituted phenylthiohydantoin which can be washed off and identified by chromatography, and the cycle can be repeated. The efficiency of each step is about 98%, which allows about 50 amino acids to be reliably determined.
Read more about this topic: Protein Sequencing
Famous quotes containing the words degradation and/or reaction:
“I suppose one has a greater sense of intellectual degradation after an interview with a doctor than from any human experience.”
—Alice James (1848–1892)
“Christianity was only a very strong and singularly well-timed Salvation Army movement that happened to receive help from an unusual and highly dramatic incident. It was a Puritan reaction in an age when, no doubt, a Puritan reaction was much wanted; but like all sudden violent reactions, it soon wanted reacting against.”
—Samuel Butler (1835–1902)