Proteomics

Dataset Information

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The MbovP280 of mycoplasma bovis immunoprecipitates with the wole cell lysates of bovine macrophage (BoMac),and then LC-MS/MS was used to identify the MbovP280-binding ligands


ABSTRACT: An IP–MS method was used to screen for MbovP280-binding proteins. Briefly, BoMac cells were cultured, harvested, and lysed in RIPA buffer supplemented with cOmplete Protease Inhibitor Cocktail (Roche, Mannheim, Germany). The whole-cell lysates (400 μg) were incubated with 10 μg of either rMbovP280 or rMbovP280∆210–269 for 1 h at 4 C. Mouse antiserum (5 μg) directed against MbovP280 was added to the lysates for 12 h at 4 C, and then 50 μl of Protein A/G Agarose Beads (Beyotime) was added for an additional 1 h. The immunoprecipitates were extensively washed with NP-40 buffer and eluted with SDS loading buffer by boiling them for 5 min. The cellular proteins that coprecipitated with rMbovP280 or rMbovP280∆210–269 were resolved with SDS-PAGE and stained with silver staining.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Cell Culture, Macrophage

SUBMITTER: Gang Zhao  

LAB HEAD: Aizhen Guo

PROVIDER: PXD022080 | Pride | 2021-09-09

REPOSITORIES: Pride

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Publications

Novel Secreted Protein of <i>Mycoplasma bovis</i> MbovP280 Induces Macrophage Apoptosis Through CRYAB.

Zhao Gang G   Zhu Xifang X   Zhang Hui H   Chen Yingyu Y   Schieck Elise E   Hu Changmin C   Chen Huanchun H   Guo Aizhen A  

Frontiers in immunology 20210215


<i>Mycoplasma bovis</i> causes important diseases and great losses on feedlots and dairy farms. However, there are only a few measures to control <i>M. bovis</i>-related diseases. As in other mycoplasma species, this is predominantly because the virulence related factors of this pathogen are largely unknown. Therefore, in this study, we aimed to identify novel virulence-related factors among the secreted proteins of <i>M. bovis</i>. Using bioinformatic tools to analyze its secreted proteins, we  ...[more]

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