%0 Journal Article %1 warscheidCharacterizationBacillusSpore2003 %A Warscheid, Bettina %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 anthracis/chemistry/isolation cereus/chemistry/isolation purification,Bacillus purification,Bacillus/chemistry/*isolation purification,Bacterial purification,to_read,Trifluoroacetic subtilis/chemistry/isolation thuringiensis/chemistry/isolation %N 20 %P 5618--5627 %R 10.1021/ac034200f %T Characterization of Bacillus Spore Species and Their Mixtures Using Postsource Decay with a Curved-Field Reflectron. %V 75 %X A strategy is proposed for the rapid identification of Bacillus spores, which relies on the selective release of a family of proteins, referred to as small, acid-soluble spore proteins (SASPs). In this work, SASPs were selectively solubilized from Bacillus spores on the MALDI sample plate by using 10\% TFA. Proteolytic digests of SASPs generated in situ from spores of B. subtilis 168, B. globigii, B. thuringiensis subs. Kurstaki HD-1, B. cereus T, and the nonpathogenic strain B. anthracis Sterne were prepared in 5-25 min by using trypsin immobilized on Agarose beads and subsequently analyzed by MALDI-TOFMS using a curved-field reflectron. Protein identification was obtained by partial sequencing of distinctive tryptic peptides from Bacillus spores via post-source decay analysis combined with genome-based database searches by Mascot Sequence Query. Various unique SASPs were identified, allowing the characterization of Bacillus species by obtaining sequence-specific information on single peptides. The applicability of this approach for the rapid identification of Bacillus species was further established by analyzing spore mixtures.

@article{warscheidCharacterizationBacillusSpore2003, abstract = {A strategy is proposed for the rapid identification of Bacillus spores, which relies on the selective release of a family of proteins, referred to as small, acid-soluble spore proteins (SASPs). In this work, SASPs were selectively solubilized from Bacillus spores on the MALDI sample plate by using 10\% TFA. Proteolytic digests of SASPs generated in situ from spores of B. subtilis 168, B. globigii, B. thuringiensis subs. Kurstaki HD-1, B. cereus T, and the nonpathogenic strain B. anthracis Sterne were prepared in 5-25 min by using trypsin immobilized on Agarose beads and subsequently analyzed by MALDI-TOFMS using a curved-field reflectron. Protein identification was obtained by partial sequencing of distinctive tryptic peptides from Bacillus spores via post-source decay analysis combined with genome-based database searches by Mascot Sequence Query. Various unique SASPs were identified, allowing the characterization of Bacillus species by obtaining sequence-specific information on single peptides. The applicability of this approach for the rapid identification of Bacillus species was further established by analyzing spore mixtures.}, added-at = {2024-05-17T13:01:35.000+0200}, address = {United States}, author = {Warscheid, Bettina and Fenselau, Catherine}, biburl = {https://www.bibsonomy.org/bibtex/2b7bb8c6da94390be4b74297c15ccddbb/warscheidlab}, doi = {10.1021/ac034200f}, interhash = {82d4467d8d11965cbb6d62296cf2f6dc}, intrahash = {b7bb8c6da94390be4b74297c15ccddbb}, 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 anthracis/chemistry/isolation 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 = {5618--5627}, pmid = {14710846}, timestamp = {2024-05-17T13:01:35.000+0200}, title = {Characterization of {{Bacillus}} Spore Species and Their Mixtures Using Postsource Decay with a Curved-Field Reflectron.}, volume = 75, year = 2003 }