title: Gel-free proteomic identification of the Bacillus subtilis insoluble spore coat protein fraction.
abstract: Species from the genus Bacillus have the ability to form endospores, dormant cellular forms that are able to survive heat and acid preservation techniques commonly used in the food industry. Resistance characteristics of spores towards various environmental stresses are in part attributed to their coat proteins. Previously, 70 proteins have been assigned to the spore coat of Bacillus subtilis using SDS-PAGE, 2-DE gel approaches, protein localization studies and genome-wide transcriptome studies. Here, we present a "gel-free" protocol that is capable of comprehensive B. subtilis spore coat protein extraction and addresses the insoluble coat fraction. Using LC-MS/MS we identified 55 proteins from the insoluble B. subtilis spore coat protein fraction, of which 21 are putative novel spore coat proteins not assigned to the spore coat until now. Identification of spore coat proteins from a B. subtilis food-spoilage isolate corroborated a generic and "applied" use of our protocol. To develop specific and sensitive spore detection and/or purification systems from food stuff or patient material, suitable protein targets can be derived from our proteomic approach. Finally, the protocol can be extended to study cross-linking among the spore coat proteins as well as for their quantification.
authors: Swammerdam Institute for Life Sciences, Department of Mass Spectrometry of Biomacromolecules, University of Amsterdam, The Netherlands. w.r.abhyankar@uva.nl
authors: Wishwas
authors: W
authors: Abhyankar
authors: Alex Ter
authors: AT
authors: Beek
authors: Henk
authors: H
authors: Dekker
authors: Remco
authors: R
authors: Kort
authors: Stanley
authors: S
authors: Brul
authors: Chris G
authors: CG
authors: de Koster
mesh_terms: D001412:Bacillus subtilis / Q000737:chemistry*; D001426:Bacterial Proteins / Q000032:analysis* / Q000302:isolation & purification; D002853:Chromatography, Liquid / Q000379:methods; D005516:Food Microbiology; D040901:Proteomics / Q000379:methods*; D012995:Solubility; D013171:Spores, Bacterial / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D001426:Bacterial Proteins; C030069:spore-specific proteins, Bacillus