Dataset Information

B. abortus 104M LC-MS/MS - A comprehensive proteogenomic study of the human Brucella vaccine strain 104 M

ABSTRACT: In the present study, we performed subcellular fractionation prior to protein extraction to reduce interference. Whole cell protein and membrane protein extracts were prepared for strain 104M. Peptide fractionation prior to analysis is an alternative method that achieves higher proteome coverage. Complexity reduction was achieved by SDS-PAGE and strong cation exchange (SCX) HPLC separation.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Brucella Abortus 104m

TISSUE(S): Cell Culture

SUBMITTER: Xiaodong Zai

LAB HEAD: Xiaodong Zai

PROVIDER: PXD005403 | Pride | 2017-05-29

REPOSITORIES: Pride

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A comprehensive proteogenomic study of the human Brucella vaccine strain 104 M.

Zai Xiaodong X Yang Qiaoling Q Liu Kun K Li Ruihua R Qian Mengying M Zhao Taoran T Li Yaohui Y Yin Ying Y Dong Dayong D Fu Ling L Li Shanhu S Xu Junjie J Chen Wei W

BMC genomics 20170523 1

<h4>Background</h4>Brucella spp. are Gram-negative, facultative intracellular pathogens that cause brucellosis in both humans and animals. The B. abortus vaccine strain 104 M is the only vaccine available in China for the prevention of brucellosis in humans. Although the B. abortus 104 M genome has been fully sequenced, the current genome annotations are not yet complete. In addition, the main mechanisms underpinning its residual toxicity and vaccine-induced immune protection have yet to be eluc ...[more]

PMID: 28535754

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Project description:Human saliva has been commonly used as protein source in in vitro microbiological and biological assays to mimic the protein pellicle formation, termed acquired salivary pellicle, that precedes microbial and cell adhesion on surfaces exposed to the oral environment. However, saliva requires previous processing to remove food debris, microorganisms, and other molecules prior its use in microbiological and biological in vitro assays. For this purpose, 0.22 μm filtration, 0.45 μm filtration, and pasteurization methods have been commonly used, but the effect of these processing methods on the proteomic profile of saliva has not been tested experimentally. Stimulated human saliva was collected from 8 healthy volunteers and submitted to the following processes: non-processing (control), 0.22 μm filtration, 0.45 μm filtration, and pasteurization. The proteomic profile of non-processed saliva was compared with 0.22 μm filtered-, 0.45 μm filtered-, and pasteurized-saliva by liquid chromatography-mass spectrometry. The effect of processed saliva in microbial adhesion was tested using bacterial and fungus species, and in biological cell behavior using HaCaT immortalized human keratinocytes. Two hundred seventy-eight proteins were identified in non-processed saliva, 54 proteins (≈19%) were exclusive. Saliva processing reduced identified proteins to 222 (≈80%) for the 0.22 μm filtered saliva, 219 (≈79%) for the 0.45 μm filtered saliva, and 201 (≈72%) for the pasteurized saliva, compared to non-processed saliva. Although there were slight differences in the protein composition, the proteomic profile showed similar molecular functions and biological processes. The different saliva processing methods did not alter microbial adhesion (ANOVA, p>0.05). Interestingly, pasteurized saliva reduced keratinocytes cell viability. Saliva processing methods tested reduced the proteomic profile diversity of saliva, but maintained similar molecular functions and biological processes mediated by remaining proteins, not interfering on microbial adhesion and cell viability, except for pasteurization, which reduced cell viability.

Dataset Information

B. abortus 104M LC-MS/MS - A comprehensive proteogenomic study of the human Brucella vaccine strain 104 M

Publications

A comprehensive proteogenomic study of the human Brucella vaccine strain 104 M.

Similar Datasets

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