Selective determination of arginine-containing and tyrosine-containing peptides using capillary electrophoresis and laser-induced fluorescence detection.
The use of two different amino acid-selective fluorogenic reagents for the derivatization of peptides is investigated. One such scheme utilizes a selective reaction of benzoin with the guanidine moiety to derivatize arginine residues occurring in a peptide. The second scheme involves the formylation of tyrosine, followed by reaction with 4-methoxy-1,2-phenylenediamine. The use of capillary electrophoresis and laser-induced fluorescence detection allows enhanced efficiencies and sensitivities to be obtained for the separations of either arginine- or tyrosine-containing peptides. A helium-cadmium laser (325 nm) is ideally suited for the laser-based detection system due to a close match of the excitation maxima of derivatized peptides from both reactions. A detection limit of 270 amol is achieved for model arginine-containing peptides, while the detection limit for model tyrosine-containing peptides is measured at 390 amol. Both derivatization reactions are found to be useful for high-sensitivity peptide mapping applications in which only the peptides containing the derivatized amino acids are detected.
Cobb
K A
KA
Department of Chemistry, Indiana University, Bloomington 47405.
Novotny
M V
MV
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001120:Arginine / Q000737:chemistry*; D002645:Chickens; D003574:Cytochrome c Group / Q000737:chemistry; D004586:Electrophoresis / Q000379:methods; D006736:Horses; D006863:Hydrogen-Ion Concentration; D007834:Lasers; D008969:Molecular Sequence Data; D009113:Muramidase / Q000737:chemistry; D009206:Myocardium / Q000201:enzymology; D010455:Peptides / Q000032:analysis*; D013050:Spectrometry, Fluorescence; D014443:Tyrosine / Q000737:chemistry*
D000596:Amino Acids; D003574:Cytochrome c Group; D010455:Peptides; D014443:Tyrosine; D001120:Arginine; D009113:Muramidase