Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

A0913 interacts with CP47 of PSII

ABSTRACT: Co-immunoprecipitation (CoIP) was performed to identify the proteins that A0913 interacts with. The thylakoid membranes were solubilized with the detergent n-Dodecyl-β-D-maltoside (DDM) before CoIP was performed using the monoclonal antibodies against the Flag tag to precipitate the target proteins. Four proteins were obtained by the CoIP as revealed by SDS-PAGE analysis. Immunoblotting with antibodies against CP47 and the Flag tag revealed that the top band in the gel was CP47 while the third band from the top down was A0913CF . Mass spectrometry revealed that these bands were CP47 of PSII.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Synechococcus Sp. (strain Atcc 27264 / Pcc 7002 / Pr-6) (agmenellum Quadruplicatum)

SUBMITTER: Zhenggao Zheng

LAB HEAD: Jindong Zhao

PROVIDER: PXD057596 | Pride | 2025-01-09

REPOSITORIES: pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	0913CF_Flag_IP_CP47.raw	Raw
	0913CF_Flag_IP_CP47.xlsx	Xlsx
	BNPAGE_0913CF_COIP.raw	Raw
	BNPAGE_0913CF_COIP.xlsx	Xlsx
	BNPAGE_WT_COIP.raw	Raw

Items per page:

1 - 5 of 8

Similar Datasets

A0913 interacts with CP47 of PSII and ApcF of PBS

Project description:Co-immunoprecipitation (CoIP) was performed to identify the proteins that A0913 interacts with. The thylakoid membranes were solubilized with the detergent n-Dodecyl-β-D-maltoside (DDM) before CoIP was performed using the monoclonal antibodies against the Flag tag to precipitate the target proteins. Four proteins were obtained by the CoIP as revealed by SDS-PAGE analysis . Immunoblotting with antibodies against CP47 and the Flag tag revealed that the top band in the gel was CP47 while the third band from the top down was A0913CF . Mass spectrometry revealed that these bands were CP47 of PSII。

2024-11-06 | PXD057430 | Pride

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

Project description: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.)

2020-08-03 | PXD019016 | Pride

SvenBldMWhiIChIPSeq

Project description:To determine the influence of BldM and WhiI on the binding of each other to their respective binding sites in the genome of Streptomyces venezuelae, ChIP-Seq experiments were carried out using polyclonal antibodies against the wild type proteins as well as anti-FLAG antibodies against FLAG-tagged proteins. Null mutants of Streptomyces venezuelae lacking WhiI or BldM were used as controls. The wild tpe strain was used as control in experiments using the anti-FLAG antibodies.

2015-05-19 | E-MTAB-2698 | biostudies-arrayexpress

Genome-wide identification of CCA1-bound genes in Arabidopsis by ChIP-Seq

Project description:LHY and CCA1 encode single MYB transcription factors, involved in circadian clock. However, direct target genes of LHY and CCA1 in a genomic scale were largely unknown. To reveal bound genes by CCA1, chimeric protein CCA1-FLAG was expressed under CCA1 promoter in cca1 lhy (CCA1pro:CCA1-FLAG/ cca1 lhy). ChIP was performed using anti-FLAG antibody (F3165; SIGMA), which was bound to Dynabeads Protein G (100-03D; Life Technologies), and ChIP DNA were analyzed by IonPGM or Illumina GAII. Chromatin immunoprecipitation was performed for CCA1-FLAG-expressing Arabidopsis. ChIP DNA was analyzed 2 types of deep sequencers (Illumina GAII and IonPGM).

2016-03-11 | E-GEOD-67903 | biostudies-arrayexpress

Heptad-specific phosphorylation of RNA polymerase II CTD

Project description:The carboxy-terminal domain (CTD) of RNA polymerase II (Pol II) consists of heptad repeats with the consensus motif Y1-S2-P3-T4-S5-P6-S7. Dynamic phosphorylation of the CTD coordinates Pol II progression through the transcription cycle. Monoclonal antibodies have been used to study in vivo the potentially phosphorylated CTD amino acids (Y1, S2, T4, S5 and S7). However, the epitopes detected by antibodies can be masked by proteins or modifications at neighbouring sites. Therefore, the effectiveness of antibodies in western blot or ChIP analysis reflects the number of accessible CTD phosphorylation marks, but not the total number of phosphorylations. Most importantly, CTD phospho-specific antibodies do not provide any heptad - (location) specific information of CTD phosphorylation. Due to these limitations, the principles and patterns of CTD phosphorylation remained elusive. Here, we use genetic and mass spectrometric approaches to directly detect and map phosphosites along the entire CTD. We confirm phosphorylation of CTD residues Y1, S2, T4, S5 and S7 in mammalian and yeast cells. Although specific phosphorylation signatures dominate, adjacent CTD repeats can be differently phosphorylated, leading to a high variation of coexisting phosphosites in mono- and di-heptad CTD repeats. Inhibition of CDK9 kinase specifically reduces S2 phosphorylation levels within the CTD.

2016-01-25 | PXD003159 | Pride

Genome-wide maps of PRR5 binding loci in Arabidopsis thaliana

Project description:PRR5 transcription factor acts in the circadian clock system. To elucidate bound genes by PRR5, Chimeric protein FLAG-PRR5-GFP, was expressed under PRR5 promoter in Col-0 (PRR5pro:FLAG-PRR5-GFP/prr5). ChIP was performed using anti-GFP antibody (ab290;Abcam), which was bound to Dynabeads Protein G (100-03D;Life technologies), and ChIP DNA and input DNA were analyzed by Illumina GA II. ChIP and input sequence reads were compared to find out PRR5 binding sites

2012-09-20 | E-GEOD-36361 | biostudies-arrayexpress

Identification of target antigens of CSF OCB-derived monoclonal antibodies

Project description:The identification of target antigens recognized by monoclonal antibodies that have been derived from oligoclonal band IgG of CSF samples from multiple sclerosis patients.

2016-06-06 | GSE72789 | GEO

Mouse ESC Nr1h2 interactome under T09 treatment

Project description:The intrinsic regulators of stem cell-based embryo models are not well studied. We show that Nr1h2 activation enables conventional ESCs to form blastoids with better implantation and higher blastocyst rates. We identified Nr1h2 transcriptional interactome by CoIP-MS using FLAG antibody to immunoprecipitate Nr1h2-FLAG and its partners in ESC overexpressing Nr1h2-FLAG.

2024-10-28 | PXD057257 | Pride

LC-MS analysis of in vitro SUMOylated SRFR1 protein.

Project description:FLAG-SRFR1 was expressed in reconstituted E. coli system (Okada el al 2009) in the presence of SUMO1 T91R SUMOylation-competent (containing diglycine, GG) or -deficient (diglycine replaced by dialanine, AA) isoforms, SCE1a, and SAE1/2. After induction of expressions, total extracts were probed with anti-FLAG antibodies. A slower migrating band than FLAG-SRFR1 was detected in the presence of only GG, but not AA forms of co- expressed SUMO1. We then subjected the slower migrating protein bands of SRFR1 expressed with SUMO1 T91R for trypsin digestion and LC-MS/MS analysis. SUMO1, but not SUMO2 or SUMO3 footprints on selective lysine residues of SRFR1 was detected in the identified peptides. Remarkably, lysine residue present in the previously predicted LK325EE SUMOylation motif was identified with SUMO1-modifications. In addition, the K229 residue of SRFR1 was also covalently modified by SUMO1 even though conserved features of a SUMOylation motif was absent for this lysine.

2022-02-17 | PXD026117 | Pride

TTC7A Localizes to Specific Regions of Chromatin

Project description:Inherited TTC7A loss of function mutations causes intestinal and immune deficiency. TTC7A is expressed in hematopoietic and epithelial cells however its cellular function remains poorly understood. In this work we provided evidence that TTC7A is an intrinsic nucleus factor. In an attempt to link the function of TTC7A in chromatin compaction, histone modifications and general transcriptional regulation we undertook to map the observed interaction of TTC7A to chromatin genome-wide by ChiP-Seq of Flag-tagged WT_TTC7A expressed in B lymphoblastoid cell lines using two anti-Flag antibodies (hereafter IP-Flag1 and IP-Flag2).

2018-10-19 | E-MTAB-5938 | biostudies-arrayexpress