MALDI Quadrupole Time-of-Flight Mass Spectrometry:  A Powerful Tool for Proteomic Research

• Published in: Analytical chemistry (Washington) 2000-05, Vol.72 (9), p.2132-2141
• Main Authors: Shevchenko, Andrej, Loboda, Alexander, Shevchenko, Anna, Ens, Werner, Standing, Kenneth G
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Hydrolysis; Indicators and Reagents; Molecular Sequence Data; Peptide Mapping; Proteins; Proteins - chemistry; Scientific imaging; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Trypsin
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac9913659

A MALDI QqTOF mass spectrometer has been used to identify proteins separated by one-dimensional or two-dimensional gel electrophoresis at the femtomole level. The high mass resolution and the high mass accuracy of this instrument in both MS and MS/MS modes allow identification of a protein either by peptide mass fingerprinting of the protein digest or from tandem mass spectra acquired by collision-induced dissociation of individual peptide precursors. A peptide mass map of the digest and tandem mass spectra of multiple peptide precursor ions can be acquired from the same sample in the course of a single experiment. Database searching and acquisition of MS and MS/MS spectra can be combined in an interactive fashion, increasing the information value of the analytical data. The approach has demonstrated its usefulness in the comprehensive characterization of protein in-gel digests, in the dissection of complex protein mixtures, and in sequencing of a low molecular weight integral membrane protein. Proteins can be identified in all types of sequence databases, including an EST database. Thus, MALDI QqTOF mass spectrometry promises to have remarkable potential for advancing proteomic research.