Solid-phase sequencing on the gas-phase sequencer.
Automated Edman degradation has been successfully used for determining the primary structure of numerous peptides and proteins. Quantitative solid-phase Edman degradation has great potential use for amino acid sequence analysis of synthetic peptides assembled on resin support by the Merrifield procedure. We report here the combined use of a modified gas-phase sequencer program and our improved reversed-phase HPLC analysis for PTH-amino acids to carry out the sequence analysis on synthesized peptide resins. This approach is far more sensitive than using glass beads on the conventional solid-phase sequencer. The peptide was assembled on copoly (styrene-1% divinylbenzene) resin beads at an initial substitution of 0.54 mmol/g. On a routine basis, 10-15 resin beads are used, and a repetitive yield of 94% is obtained: as few as 4 beads can be successfully sequenced. The HPLC PTH-amino acid analysis is sensitive down to subpicomole quantities. This procedure offers a sensitive and rapid analytical tool for checking the purity of peptides as they are being assembled on solid support.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D009320:Atrial Natriuretic Factor / Q000032:analysis; D001322:Autoanalysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D008832:Microchemistry; D010669:Phenylthiohydantoin / Q000031:analogs & derivatives / Q000032:analysis; D012116:Resins, Plant
D000596:Amino Acids; D012116:Resins, Plant; D010669:Phenylthiohydantoin; D009320:Atrial Natriuretic Factor