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Identification of marker proteins for Bacillus anthracis using MALDI‐TOF MS and ion trap MS/MS after direct extraction or electrophoretic separation
Author(s) -
Stump Michael J.,
Black Gavin,
Fox Alvin,
Fox Karen F.,
Turick Charles E.,
Matthews Michael
Publication year - 2005
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200500143
Subject(s) - chromatography , chemistry , bacillus anthracis , electrophoresis , extraction (chemistry) , ion trap , mass spectrometry , matrix assisted laser desorption/ionization , trap (plumbing) , time of flight mass spectrometry , ion , biology , bacteria , ionization , genetics , environmental engineering , engineering , organic chemistry , adsorption , desorption
Direct extraction of bacterial vegetative cells or spores followed by matrix‐assisted laser desorption ionization/time of flight mass spectrometry (MALDI TOF MS) has become popular for bacterial identification, since it is simple to perform and mass spectra are readily interpreted. However, only high‐abundance proteins that are of low mass and ionize readily are observed. In the case of B. anthracis spores, small acid‐soluble spore proteins (SASPs) have been the most widely studied. Additional information can be obtained using tandem mass spectrometry (MS‐MS) to confirm the identity of proteins by sequencing. This is most readily accomplished using ion trap (IT) MS‐MS. However, enzymatic digestion of these proteins is needed to generate peptides that are within the mass range of the ion trap. The use of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE), or other forms of electrophoresis, allows one to focus on specific proteins of interest ( e. g. the high mass exosporium glycoproteins BclA and BclB) that provide additional species‐ and strain‐specific discrimination.