Title
Gel-free proteomic identification of the Bacillus subtilis insoluble spore coat protein fraction
Author
Abhyankar, W.
Beek, A.T.
Dekker, H.
Kort, R.
Brul, S.
de Koster, C.G.
Publication year
2011
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. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Subject
Life
MSB - Microbiology and Systems Biology
EELS - Earth, Environmental and Life Sciences
Healthy Living
Bacillus
Microbiology
MS
Spore coat
coat protein
article
Bacillus subtilis
endospore-forming rods and cocci
environmental stress
food industry
food spoilage
liquid chromatography
mass spectrometry
nonhuman
polyacrylamide gel electrophoresis
priority journal
protein analysis
protein localization
protein targeting
signal transduction
solubility
two dimensional gel electrophoresis
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DOI
https://doi.org/10.1002/pmic.201100003
TNO identifier
445990
ISSN
1615-9853
Source
Proteomics, 11 (23), 4541-4550
Document type
article