title: Reinforcement of Bacillus subtilis spores by cross-linking of outer coat proteins during maturation.
abstract: Resistance characteristics of bacterial endospores towards various environmental stresses such as chemicals and heat are in part attributed to their coat proteins. Heat resistance is developed in a late stage of sporulation and during maturation of released spores. Using our gel-free proteomic approach and LC-FT-ICR-MS/MS analysis we have monitored the efficiency of the tryptic digestion of proteins in the coat during spore maturation over a period of eight days, using metabolically (15)N labeled mature spores as reference. The results showed that during spore maturation the loss of digestion efficiency of outer coat and crust proteins synchronized with the increase in heat resistance. This implicates that spore maturation involves chemical cross-linking of outer coat and crust layer proteins leaving the inner coat layer proteins unmodified. It appears that digestion efficiencies of spore surface proteins can be linked to their location within the coat and crust layers. We also attempted to study a possible link between spore maturation and the observed heterogeneity in spore germination.
authors.affiliation: Mass Spectrometry of Biomacromolecules, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands; Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Wishwas
authors.initials: W
authors.lastname: Abhyankar
authors.affiliation: Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Rachna
authors.initials: R
authors.lastname: Pandey
authors.affiliation: Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Alexander
authors.initials: A
authors.lastname: Ter Beek
authors.affiliation: Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Stanley
authors.initials: S
authors.lastname: Brul
authors.affiliation: Mass Spectrometry of Biomacromolecules, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Leo J
authors.initials: LJ
authors.lastname: de Koning
authors.affiliation: Mass Spectrometry of Biomacromolecules, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands. Electronic address: C.G.deKoster@uva.nl.
authors.forename: Chris G
authors.initials: CG
authors.lastname: de Koster
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D002853:Chromatography, Liquid; D003432:Cross-Linking Reagents; D005516:Food Microbiology; D006358:Hot Temperature; D040901:Proteomics; D013171:Spores, Bacterial; D053719:Tandem Mass Spectrometry; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; C030069:spore-specific proteins, Bacillus
keywords: Bacillus subtilis; Germination; LC-FT-ICR-MS/MS; Protein cross-linking; Spore coat; Spore maturation; Thermal resistance