Rapid tryptic mapping using enzymatically active mass spectrometer probe tips.
A method has been developed for rapid, sensitive, and accurate tryptic mapping of polypeptides using matrix-assisted laser desorption/ionization time-of-flight mass analysis. The technique utilizes mass spectrometer probe tips which have been activated through the covalent immobilization of trypsin. The enzymatically active probe tips were used for the tryptic mapping of chicken egg lysozyme and the results compared with those obtained using either free trypsin or agarose-immobilized trypsin. A significant increase in the overall sensitivity of the process was observed using the active probe tips, as well as the production of more characteristic proteolytic fragments and the elimination of background signals due to the autolysis of the trypsin. Further, probe tip digestions were found to be rapid and convenient.
Dogruel
D
D
Department of Chemistry and Biochemistry, Arizona State University, Tempe 85287-1604, USA.
Williams
P
P
Nelson
R W
RW
D000595:Amino Acid Sequence; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D014357:Trypsin / Q000494:pharmacology
D014357:Trypsin