Mass spectrometric sequencing of peptides and proteins

Abstract

-Mass spectrometry had been demonstrated to be a useful technique for the determination of the amino acid sequence in oligopeptides. The extension to the sequencing of polypeptides and proteins is a complex problem involving chemical degradation and conversion techniques, separations, collection of the mass spectral information and the interpretation thereof. Partial acid hydrolyzates of primary degradation peptides (consisting of about 50 amino acids) of proteins are converted to O-TMS-polyamino alcohols which are analyzed by gas chromatographic mass spectrometry to identify all the peptides formed in the hydrolysis. From this information the original structure is then reassembled. The method is illustrated with a cyclic peptide, with the N-terminal sequence of A repressor and the structure of the sweet protein monelln.

Fast atom bombardment and tandem mass spectrometry for sequencing peptides and polyamino alcohols

“…Biemann and co-workers have developed another approach based on chemical reduction of peptides to their corresponding polyamino alcohols (4). They have carefully evaluated the reduction and coupled it with GC/MS for analysis of the product mixtures which result from chemical reduction of a mixture of peptides from a digest (5)(6)(7)(8)(9)(10)(11)(12)(13).…”

Analytical techniques for trace organic compounds - II. Detectors for gas chromatography

“…The principles of operation of these mass spectrometers have been discussed by Biemann in another IUPAC paper of this series (ref. 27) and will not be repeated here.…”