A novel ion trap time-of-flight hybrid mass spectrometer (qIT-TOF MS) has been applied for peptide sequencing in proteolytic digests generated from spore mixtures of Bacilli. The method of on-probe solubilization and in situ proteolytic digestion of small, acid-soluble spore proteins has been recently developed in our laboratory, and microorganism identification in less than 20 min was accomplished. In this study, tryptic peptides were generated in situ from complex spore mixtures of B. subtilis 168, B. globigii, B. thuringiensis subs. Kurstaki, and B. cereus T, respectively. MALDI analysis of bacterial peptides generated was performed with an average mass resolving power of 6200 and a mass accuracy of up to 10 ppm using a trap-TOF tandem configuration. Precursor ions of interest were usually selected and stored in the quadrupole ion trap with their complete isotope distribution by choosing a window of +/- 2 Da. Sequence-specific information on isolated protonated peptides was gained via tandem MS experiments with an average mass resolving power of 4450 for product ion analysis, and protein and bacterial sources were identified by database searching.
%0 Journal Article
%1 warscheidMALDIAnalysisBacilli2003
%A Warscheid, Bettina
%A Jackson, Kathryn
%A Sutton, Chris
%A Fenselau, Catherine
%C United States
%D 2003
%J Analytical chemistry
%K & Acid Acid/chemistry,Trypsin/chemistry/metabolism Amino Bacterial/chemistry/*isolation Data,Molecular Desorption-Ionization/*methods,Spores Fragments/analysis,Spectrometry Laser Mass Matrix-Assisted Protein,Molecular Proteins/chemistry/metabolism,Databases Sequence Sequence,Bacillus Weight,Peptide cereus/chemistry/isolation purification,Bacillus purification,Bacillus/chemistry/*isolation purification,Bacterial purification,to_read,Trifluoroacetic subtilis/chemistry/isolation thuringiensis/chemistry/isolation
%N 20
%P 5608--5617
%R 10.1021/ac0344081
%T MALDI Analysis of Bacilli in Spore Mixtures by Applying a Quadrupole Ion Trap Time-of-Flight Tandem Mass Spectrometer.
%V 75
%X A novel ion trap time-of-flight hybrid mass spectrometer (qIT-TOF MS) has been applied for peptide sequencing in proteolytic digests generated from spore mixtures of Bacilli. The method of on-probe solubilization and in situ proteolytic digestion of small, acid-soluble spore proteins has been recently developed in our laboratory, and microorganism identification in less than 20 min was accomplished. In this study, tryptic peptides were generated in situ from complex spore mixtures of B. subtilis 168, B. globigii, B. thuringiensis subs. Kurstaki, and B. cereus T, respectively. MALDI analysis of bacterial peptides generated was performed with an average mass resolving power of 6200 and a mass accuracy of up to 10 ppm using a trap-TOF tandem configuration. Precursor ions of interest were usually selected and stored in the quadrupole ion trap with their complete isotope distribution by choosing a window of +/- 2 Da. Sequence-specific information on isolated protonated peptides was gained via tandem MS experiments with an average mass resolving power of 4450 for product ion analysis, and protein and bacterial sources were identified by database searching.
@article{warscheidMALDIAnalysisBacilli2003,
abstract = {A novel ion trap time-of-flight hybrid mass spectrometer (qIT-TOF MS) has been applied for peptide sequencing in proteolytic digests generated from spore mixtures of Bacilli. The method of on-probe solubilization and in situ proteolytic digestion of small, acid-soluble spore proteins has been recently developed in our laboratory, and microorganism identification in less than 20 min was accomplished. In this study, tryptic peptides were generated in situ from complex spore mixtures of B. subtilis 168, B. globigii, B. thuringiensis subs. Kurstaki, and B. cereus T, respectively. MALDI analysis of bacterial peptides generated was performed with an average mass resolving power of 6200 and a mass accuracy of up to 10 ppm using a trap-TOF tandem configuration. Precursor ions of interest were usually selected and stored in the quadrupole ion trap with their complete isotope distribution by choosing a window of +/- 2 Da. Sequence-specific information on isolated protonated peptides was gained via tandem MS experiments with an average mass resolving power of 4450 for product ion analysis, and protein and bacterial sources were identified by database searching.},
added-at = {2024-05-17T13:01:35.000+0200},
address = {United States},
author = {Warscheid, Bettina and Jackson, Kathryn and Sutton, Chris and Fenselau, Catherine},
biburl = {https://www.bibsonomy.org/bibtex/2a57652b7d85548292d1f19091c44e537/warscheidlab},
doi = {10.1021/ac0344081},
interhash = {80e55d97dae4e4fc6224e6329cfc3a2f},
intrahash = {a57652b7d85548292d1f19091c44e537},
issn = {0003-2700},
journal = {Analytical chemistry},
keywords = {& Acid Acid/chemistry,Trypsin/chemistry/metabolism Amino Bacterial/chemistry/*isolation Data,Molecular Desorption-Ionization/*methods,Spores Fragments/analysis,Spectrometry Laser Mass Matrix-Assisted Protein,Molecular Proteins/chemistry/metabolism,Databases Sequence Sequence,Bacillus Weight,Peptide cereus/chemistry/isolation purification,Bacillus purification,Bacillus/chemistry/*isolation purification,Bacterial purification,to_read,Trifluoroacetic subtilis/chemistry/isolation thuringiensis/chemistry/isolation},
langid = {english},
month = oct,
number = 20,
pages = {5608--5617},
pmid = {14710845},
timestamp = {2024-05-17T13:01:35.000+0200},
title = {{{MALDI}} Analysis of {{Bacilli}} in Spore Mixtures by Applying a Quadrupole Ion Trap Time-of-Flight Tandem Mass Spectrometer.},
volume = 75,
year = 2003
}