Proteomics

Dataset Information

0

‘One-pot’ sample processing method for proteome-wide analysis of microbial cells and spores


ABSTRACT: Bacterial endospores, the transmissible forms of pathogenic bacilli and clostridia, are heterogeneous multilayered structures composed of proteins. These proteins protect the spores against variety of stresses, thus helping spore survival, and assist in germination, by interacting with the environment to form vegetative cells. Owing to the complexity, insolubility, and dynamic nature of spore proteins, it has been difficult to obtain their comprehensive protein profiles. The intact spores of Bacillus subtilis, Bacillus cereus, and Peptoclostridium difficile and their vegetative counterparts were disrupted by bead-beating in 6M urea under reductive conditions. The heterogeneous mixture was then double-digested with LysC and trypsin. Next, the peptide mixture was pre-fractionated with Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) followed by reverse phase LC-FT-MS analysis of the fractions. ‘One-pot’ method is a simple, robust method that yields identification of >1000 proteins with high confidence, across all spore layers from Bacillus subtilis, Bacillus cereus, and Peptoclostridium difficile. This method can be employed for proteome-wide analysis of non-spore-forming as well as spore-forming pathogens. Analysis of spore protein profile will help to understand the sporulation and germination processes and to distinguish immunogenic protein markers.

INSTRUMENT(S): apex ultra

ORGANISM(S): Peptoclostridium Difficile (strain 630) (clostridium Difficile) Bacillus Subtilis Py79 Bacillus Cereus (strain Atcc 14579 / Dsm 31)

SUBMITTER: Bhagyashree Swarge  

LAB HEAD: Prof. Dr. Chris G. De Koster

PROVIDER: PXD008242 | Pride | 2018-07-05

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
CW2808.baf Other
CW2808Semi-TRP.xml Xml
CW2808TRP.xml Xml
CW2808_RE2_01_2649.mgf Mgf
CW2809.baf Other
Items per page:
1 - 5 of 119
altmetric image

Publications

"One-Pot" Sample Processing Method for Proteome-Wide Analysis of Microbial Cells and Spores.

Swarge Bhagyashree Nandakishor BN   Roseboom Winfried W   Zheng Linli L   Abhyankar Wishwas R WR   Brul Stanley S   de Koster Chris G CG   de Koning Leo J LJ  

Proteomics. Clinical applications 20180416 5


<h4>Purpose</h4>Bacterial endospores, the transmissible forms of pathogenic bacilli and clostridia, are heterogeneous multilayered structures composed of proteins. These proteins protect the spores against a variety of stresses, thus helping spore survival, and assist in germination, by interacting with the environment to form vegetative cells. Owing to the complexity, insolubility, and dynamic nature of spore proteins, it has been difficult to obtain their comprehensive protein profiles.<h4>Exp  ...[more]

Similar Datasets

2018-05-01 | PXD009235 | Pride
2020-05-26 | PXD012030 | Pride
2015-01-26 | E-GEOD-65259 | biostudies-arrayexpress
2018-07-05 | PXD008592 | Pride
2018-10-26 | PXD003569 | Pride
2021-05-05 | GSE173854 | GEO
2019-10-16 | PXD010359 | Pride
2024-05-27 | PXD052347 | Pride
2014-08-14 | E-GEOD-60412 | biostudies-arrayexpress
2012-06-12 | E-GEOD-38619 | biostudies-arrayexpress