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Protein identification based on matrix assisted laser desorption/ionization‐post source decay‐mass spectrometry
Author(s) -
Gevaert Kris,
Demol Hans,
Martens Lennart,
Hoorelbeke Bart,
Puype Magda,
Goethals Marc,
Damme Jozef Van,
Boeck Stefaan De,
Vandekerckhove Joël
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200105)22:9<1645::aid-elps1645>3.0.co;2-z
Subject(s) - peptide mass fingerprinting , mass spectrometry , matrix assisted laser desorption/ionization , protein mass spectrometry , proteome , chemistry , peptide , mass spectrum , bottom up proteomics , sample preparation in mass spectrometry , chromatography , ionization , isobaric labeling , surface enhanced laser desorption/ionization , fragmentation (computing) , analytical chemistry (journal) , maldi imaging , proteomics , desorption , tandem mass spectrometry , electrospray ionization , biochemistry , biology , ion , ecology , organic chemistry , adsorption , gene
Due to its very short analysis time, its high sensitivity and ease of automation, matrix‐assisted laser desorption/ionization (MALDI)‐peptide mass fingerprinting has become the preferred method for identifying proteins of which the sequences are available in databases. However, many protein samples cannot be unambiguously identified by exclusively using their peptide mass fingerprints ( e.g., protein mixtures, heavily post‐translationally modified proteins and small proteins). In these cases, additional sequence information is needed and one of the obvious choices when working with MALDI‐mass spectrometry (MS) is to choose for post source decay (PSD) analysis on selected peptides. This can be performed on the same sample which is used for peptide mass fingerprinting. Although in this type of peptide analysis, fragmentation yields are very low and PSD spectra are often very difficult to interpret manually, we here report upon our five years of experience with the use of PSD spectra for protein identification in sequence (protein or expressed sequence tag (EST)) databases. The combination of peptide mass fingerprinting and PSD and analysis described here generally leads to unambiguous protein identification in the amount of material range generally encountered in most proteome studies.