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Labeling elastase digests with TMT: Informational gain by identification of poorly detectable peptides with MALDI‐TOF/TOF mass spectrometry
Author(s) -
Baeumlisberger Dominic,
Arrey Tabiwang N.,
Rietschel Benjamin,
Rohmer Marion,
Papasotiriou Dimitrios G.,
Mueller Benjamin,
Beckhaus Tobias,
Karas Michael
Publication year - 2010
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201000288
Subject(s) - chemistry , tandem mass tag , isobaric labeling , peptide , mass spectrometry , chromatography , protein mass spectrometry , tandem mass spectrometry , bottom up proteomics , peptide mass fingerprinting , matrix assisted laser desorption/ionization , protease , electrospray ionization , peptide sequence , sample preparation in mass spectrometry , biochemistry , proteomics , enzyme , quantitative proteomics , gene , organic chemistry , adsorption , desorption
The applicability of the less specific protease elastase for the identification of membrane and cytosolic proteins has already been demonstrated. MALDI as ionization technique particularly favors the detection of basic and to a lesser extent of weakly acidic peptides, whereas neutral peptides often remain undetected. Moreover, peptides below 700 Da are routinely excluded. In the following study, the advantage of additional information gained from tandem mass tag zero labeled peptides and the resultant increase in sequence coverage was evaluated. Through derivatization with tandem mass tag reagents, peptide measurement within the standard mass range of the MALDI reflector mode is achievable due to the mass increase. Compared to the unlabeled sample, peptides exhibiting relatively low molecular masses, p I values or higher hydrophobicity could be identified.