Proteomics

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Overcoming off-targets: Assessing western blot signals for Bcnt/Cfdp1, a tentative component of the chromatin remodeling complex

ABSTRACT: The mammalian Bucentaur/Craniofacial developmental protein 1 (Bcnt/Cfdp1) is a tentative component of the Srcap (SNF2-Related CBP Activator Protein) chromatin remodeling complex, but little is known about its properties, partly because few antibodies are available to detect the endogenous protein.　We evaluated anti-Bcnt/Cfdp1 antibodies, which were generated against either of two unrelated immuno-gens, BCNT-C domain, or mouse N-terminal peptide. As a result, we assigned the signal of the endogenous mBcnt/Cfdp1 as a doublet band of ~45-kDa using western blot analysis. LC-MS/MS analysis of the corresponding doublet to the Flag-tagged mouse Bcnt/Cfdp1 exhibited that the upper band was much more phosphorylated than the lower band having preferential Ser phosphorylation in the DWESF motif in the BCNT-C domain. These features are very similar to those of His-tagged hBCNT/CFDP1, as previously reported(Biosci Rep. 2015; 35, pii: e00228.)

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture

SUBMITTER: Takehiro Suzuki  

LAB HEAD: Naoshi Dohmae

PROVIDER: PXD019016 | Pride | 2020-08-03

REPOSITORIES: Pride

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Dataset's files

Source:
Action DRS
BCNT_API.mzML Mzml
BCNT_API.mzid.gz Mzid
BCNT_API.pride.mgf.gz Mgf
BCNT_AspN.mzML Mzml
BCNT_AspN.mzid.gz Mzid
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Publications

Overcoming off-targets: assessing Western blot signals for Bcnt/Cfdp1, a tentative component of the chromatin remodeling complex.

Iwashita Shintaro S   Suzuki Takehiro T   Kiriyama Yoshimitsu Y   Dohmae Naoshi N   Ohoka Yoshiharu Y   Song Si-Young SY   Nakashima Kentaro K  

Bioscience reports 20200601 6

The Bucentaur (BCNT) protein family is characterized by a conserved amino acid sequence at the C-terminus (BCNT-C domain) and plays an essential role in gene expression and chromosomal maintenance in yeast and Drosophila. The mammalian Bucentaur/Craniofacial developmental protein 1 (Bcnt/Cfdp1) is also a tentative component of the SNF2-related CBP activator protein (Srcap) chromatin remodeling complex, but little is known about its properties, partly because few antibodies are available to exami  ...[more]

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