Separation of peptides on the basis of the difference in positive charge: simultaneous isolation of C-terminal and blocked N-terminal peptides from tryptic digests

Abstract

The microscale separation of peptides based on the difference in positive charge was examined with tryptic digests of apomyoglobin and calmodulin. By this separation method, C-terminal and blocked N-terminal peptides could be selectively isolated in the same fraction without any chemical modifications. Separated peptides, including internal peptides, were further purified by reversed phase high performance liquid chromatography, and the purified peptides could be directly subjected to sequence and amino acid analyses. The N-terminal peptides of calcium-activated neutral protease were successfully isolated by this method.