Preparation of carboxyl-terminal tryptic peptides from proteins by cleavage at arginine.
A method has been developed for selectively preparing the carboxyl-terminal tryptic peptide of proteins by cleavage at arginyl residues. The succinylated protein is digested with trypsin and the peptides produced are maleylated. Maleylated peptides are then submitted to cation-exchange chromatography in urea at low pH and ionic strength. Arginine-containing peptides are retained by the resin. The carboxyl-terminal peptide emerges unretarded and in pure form. This method has been applied to four proteins of known sequence. Yields as high as 88% have been obtained.
D000596:Amino Acids / Q000032:analysis; D001120:Arginine / Q000032:analysis; D002852:Chromatography, Ion Exchange; D008722:Methods; D010455:Peptides / Q000032:analysis / Q000138:chemical synthesis* / Q000302:isolation & purification; D011506:Proteins; D014357:Trypsin
D000596:Amino Acids; D010455:Peptides; D011506:Proteins; D001120:Arginine; D014357:Trypsin