This communication presents a high-performance liquid chromatography system which can be used for analytical peptide mapping as well as for preparative pcptide separations. The method uses a macroreticular anion-exchange resin with an organic solvent elution system, and the eluates are monitored directly by uv absorption. Complex pcptide mixtures derived from partial hydrolysis of proteins are effectively resolved within 60 min with high reproducibility, and the eluted peptides can be analyzed for amino acid composition without any desalting processes.
"Peptide mapping" is a valuable analytical technique for identification, characterization and, more often, for comparison of proteins on structural basis. In this technique, proteins are first cleaved with proteolytic enzymes such as trypsin or with specific chemical reagents such as cyanogen bromide (I), and the resulting peptide mixtures are then fractionated by means of chromatography and/or electrophoresis of various types (2-7). Since the cleavage of protein is usually reproducible under controlled conditions, success of the mapping experiments depends mainly on the latter separation method of peptides.
The separation of peptides is also an essential, but often time-consuming step in protein sequence determination.
Recent advances of Edman chemistry and develop-' Requests for reprints should be addressed to this author.