Dataset Information

Temperature-dependent Proteome Study of Thermoanaerobacter tengcongensis

ABSTRACT: Comprehensive and quantitative information of the thermophile proteome is an important source for understanding of the survival mechanism under high growth temperature. Thermoanaerobacter tengcongensis (T. tengcongensis), a typical anaerobic thermophilic eubacterium, was selected to quantitatively evaluate its protein abundance changes in response to four different temperatures. Thermoanaerobacter tengcongensis proteins were trypsine digested, separated with high-pH RP, and identified with MS/MS analysis. The raw MS/MS data were converted into MGF format by Proteome Discoverer 1.2 (Thermo Fisher Scientific, Waltham, MA, USA). And the exported MGF files were searched by Mascot 2.3.02 (Matrix Science, Boston, MA, USA) against the database with all 2588 predicted proteins in T. tengcongensis downloaded from NCBI (NCBI reference sequence: NC_003869.1). An automatic decoy database search was performed. Several parameters in Mascot were set for peptide searching, tolerance of one missed cleavage of trypsin, Carbamidomethyl (C),iTRAQ8plex (K) and iTRAQ8plex (N-term) as fixed modification, iTRAQ8plex (Y),Oxidation (M) as variable modification. The precursor mass tolerance was 10 ppm, and the product ion tolerance was 0.02 Da.

INSTRUMENT(S): LTQ Orbitrap, LTQ Orbitrap Velos, instrument model

ORGANISM(S): Thermoanaerobacter Tengcongensis (strain Dsm 15242 / Jcm 11007 / Nbrc 100824 / Mb4) (caldanaerobacter Subterraneus Subsp. Tengcongensis)

TISSUE(S): Whole Body

SUBMITTER: Bo Wen

PROVIDER: PXD000264 | Pride | 2013-08-13

REPOSITORIES: Pride

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Publications

Quantitative proteomics reveals the temperature-dependent proteins encoded by a series of cluster genes in thermoanaerobacter tengcongensis.

Chen Zhen Z Wen Bo B Wang Quanhui Q Tong Wei W Guo Jiao J Bai Xue X Zhao Jingjing J Sun Yao Y Tang Qi Q Lin Zhilong Z Lin Liang L Liu Siqi S

Molecular & cellular proteomics : MCP 20130512 8

Comprehensive and quantitative information of the thermophile proteome is an important source for understanding of the survival mechanism under high growth temperature. Thermoanaerobacter tengcongensis (T. tengcongensis), a typical anaerobic thermophilic eubacterium, was selected to quantitatively evaluate its protein abundance changes in response to four different temperatures. With optimized procedures of isobaric tags for relative and absolute quantitation quantitative proteomics (iTRAQ), suc ...[more]

PMID: 23665590

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Project description:To identify and quantify protein profiles from peripheral blood mononuclear cells (PBMC) of osteopetrosis patients with isobaric Tagging for Relative and Absolute protein Quantification (iTRAQ) based proteomic technology and to find differentially expressed proteins in osteopetrosis. Total protein was extracted from PBMC samples of 6 osteopetrosis patients and 9 health donors. For the osteopetrosis group and the control group, aliquots of protein from each subject were pooled respectively. One hundred micrograms of protein from each corresponding group pool protein was digested with trypsin, and labeled according to the iTRAQ protocol. The labeled digests were then combined into one sample mixture and subjected to LC-MS/MS analysis. Identification and relative quantification of proteins based on the ratio of peak areas from MS/MS spectra, and the m/z of 114 (osteopetrosis) and 115 (control) were involved in this research. The MS data were processed using Proteome Discoverer Software (Version 1.2.0.208) to generate the peak list. Mascot 2.3.02 was used for the protein identification, and the parameters as follows: Type of search: MS/MS Ion search; Enzyme: Trypsin; Fragment mass tolerance: ±0.1 Da; Mass values: Monoisotopic; Variable modifications: Gln->pyro-Glu (N-term Q), Oxidation (M), iTRAQ8plex(Y); Peptide mass tolerance: 0.05 Da; Instrument type: Defult; Max missed cleavages: 1; Fixed modifications: arbamidomethyl (C), iTRAQ8plex (N-term), iTRAQ8plex (K); Protein Mass: Unrestricted; Other parameters: default; Database, IPI_human v3.87 (91464 sequences). The search results were further filtered with the parameters as follows: for protein identification, significance threshold: P<0.05 (with 95% confidence), ion score or expected cutoff < 0.05 (with 95% confidence); for protein quantitation, protein ratio type: weighted; minimum precursor charge: 1; minimum peptides: 2 (unique peptides); median intensities: normalization (removed outliers automatically). Proteins with 1.5 fold or more change between osteopetrosis group and the control group were determined as differentially expressed proteins.

Dataset Information

Temperature-dependent Proteome Study of Thermoanaerobacter tengcongensis

Publications

Quantitative proteomics reveals the temperature-dependent proteins encoded by a series of cluster genes in thermoanaerobacter tengcongensis.

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