Atmospheric pressure matrix-assisted laser desorption/ionization ion trap mass spectrometry of sulfonic acid derivatized tryptic peptides.
Atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI) and ion trap mass spectrometry have been used to study the fragmentation behavior of native peptides and peptide derivatives prepared for de novo sequencing applications. Sulfonic acid derivatized peptides were observed to fragment more extensively and up to 28 times more efficiently than the corresponding native peptides. Tandem mass spectra of native peptides containing aspartic or glutamic acids are dominated by cleavage on the C-terminal side of the acidic residues. This significantly limits the amount of sequence information that can be derived from those compounds. The MS/MS spectra of native tryptic peptides containing oxidized Met residues show extensive loss of CH(3)SOH and little sequence-specific fragmentation. On the other hand, the tandem mass spectra of derivatized peptides containing Asp, Glu and oxidized Met show much more uniform fragmentation along the peptide backbone. The AP-MALDI tandem mass spectra of some derivatized peptides were shown to be qualitatively very similar to the corresponding vacuum MALDI postsource decay mass spectra, which were obtained on a reflector time-of-flight instrument. However, the ion trap mass spectrometer offers several advantages for peptide sequencing relative to current reflector time-of-flight instruments including improved product ion mass measurement accuracy, improved precursor ion selection and MS(n). These latter capabilities were demonstrated with solution digests of model proteins and with in-gel digests of 2D-gel separated proteins.
Keough
T
T
The Procter and Gamble Company, Miami Valley Laboratories, PO Box 538707, Cincinnati, OH 45253-8707, USA. keough.tw@pg.com
Lacey
M P
MP
Strife
R J
RJ
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000032:analysis; D001274:Atmospheric Pressure; D015180:Electrophoresis, Gel, Two-Dimensional; D005340:Fibrinogen / Q000032:analysis / Q000737:chemistry; D018698:Glutamic Acid / Q000032:analysis; D007477:Ions; D008715:Methionine / Q000032:analysis / Q000378:metabolism; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry; D011392:Proline / Q000032:analysis; D015203:Reproducibility of Results; D020539:Sequence Analysis, Protein / Q000295:instrumentation / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013451:Sulfonic Acids / Q000737:chemistry*
D007477:Ions; D010446:Peptide Fragments; D013451:Sulfonic Acids; D001224:Aspartic Acid; D018698:Glutamic Acid; D005340:Fibrinogen; D011392:Proline; D008715:Methionine