Identification and characterization of posttranslational modifications of proteins by MALDI ion trap mass spectrometry.
Matrix-assisted laser desorption/ionization (MALDI) ion trap mass spectrometry is shown to be a powerful tool for the elucidation of protein modifications. Low-energy covalent bonds that originate from certain posttranslational modifications dissociate preferentially to produce characteristic mass spectrometric signatures that prove useful for the accurate, confident identification and characterization of such modifications. Because the MALDI ion trap is an authentic tandem mass spectrometer, it proves feasible to acquire secondary information to test hypotheses as to the nature and site of the putative modifications--further increasing the reliability of the tool. The method combines the advantageous features of MALDI (i.e., the ability to measure the same sample repeatedly, to measure unfractionated complex mixtures without the need for sample cleaning, and to determine peptide mixtures with subpicomole sensitivity) with the ease and the speed of the ion trap measurement. We demonstrate how the unique properties of MALDI ion trap MS can be used to address problems involving the determination of both native posttranslational modifications of proteins (e.g., disulfide mapping, glycosylation determination, and phosphorylation determination) and non-native chemical modifications of proteins (e.g., methionine oxidation and photo-cross-linking of proteins with DNA).
Qin
J
J
Rockefeller University, New York, New York 10021, USA.
Chait
B T
BT
D003432:Cross-Linking Reagents / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry; D006031:Glycosylation; D008715:Methionine / Q000378:metabolism; D010084:Oxidation-Reduction; D010449:Peptide Mapping; D010766:Phosphorylation; D010777:Photochemistry; D011499:Protein Processing, Post-Translational; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D003432:Cross-Linking Reagents; D004220:Disulfides; D008715:Methionine