Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Isolation and MS characterisation of recombinant human septin octameric complexes containing distinct SEPT9 isoforms

ABSTRACT: Native human septins form heterooctameric complexes containing different SEPT9 isoforms. To enable studies of SEPT9 function, we established a protocol for the isolation of recombinant human octamers harboring distinct SEPT9 isoforms. Mass spectrometry analysis is used to assess the purity, integrity and stoichiometry of septins in the purified complex.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Luc Camoin

LAB HEAD: Marseille Proteomics

PROVIDER: PXD023857 | Pride | 2021-07-27

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

RNA-seq analysis of Clostridium beijerinckii DSM 6423 at early exponential, acetogenic, and solventogenic growth phase on rhamnose compared to glucose

Project description:This experiment aim was to characterize the catabolism of L-rhamnose of Clostridium beijerinckii DSM 6423 by transcriptomic analysis, generating new insights and knowledge on utilization of L-rhamnose for production of chemicals, including Isopropanol, Butanol, Ethanol (IBE) and 1,2-propandiol. These analysis on cultures grown on L-rhamnose compared to D-glucose grown cultures showed upregulation of the L-rhamnose-related clusters and genes, and lower expression of the solventogenic genes, which was reflected in the products formed.

2019-01-14 | E-MTAB-7487 | biostudies-arrayexpress

Mapping the SLP76 interactome in T cells lacking each of the GRB2-family adaptors reveals molecular plasticity of the TCR signaling pathway

Project description:The propagation of intracellular T cell receptor (TCR) signals involved various adapter proteins that are important molecular switches to connect proteins together. The global characterization of changes in protein-protein interactions following genetic perturbations is critical to understand the reorganization of protein networks associated with the observed phenotypes. Here, by combining genome editing techniques and AP-MS analysis we determined and quantified the molecular reorganization of the SLP76 interactome resulting from the inactivation of each of the three GRB2-family adaptors. Our data define disruption context-specific and time-dependent networks that form around SLP76 after TCR stimulation, and selectively preserved or highly affected protein complexes that regulate T cell activation.

2023-03-13 | PXD038891 | Pride

Searching for new members of the SLX4 complex and for SLX4-complex members that are SUMOylated in vitro

Project description:The project was initially aimed at identifying new members of the SLX4 complex as well as members of the complex that are SUMOylated in vitro. The project was also aimed at assessing whether SUMOylation may change the compositon of the complex by reducing complex association of SUMOylated partners. YFP-SLX4 complexes were immunopurified from Hela Flp-In TRex cells producing full length YFP-SLX4 using a GFP nanobody. The YFP-SLX4 pull down was used in an ex vivo/in vitro SUMO-ligation assay as described in Guervilly et al. 2015. After the SUMO-ligation reaction, beads were washed 5 times with 50 mM Tris-HCl [pH 8.0] buffer, allowing for the removal of SUMOylated proteins that associate less efficiently with SLX4, or other members of the complex. SLX4 and proteins remaining associated with SLX4 after the successive washes were eluted directly in NuPAGE LDS sample buffer (Invitrogen) for 5 min at 95˚C.

2020-03-05 | PXD012425 | Pride

The RHOA-specific GTPase activating protein ARHGAP45 controls chemokine-driven migration of naïve T and B cells and of hematopoietic progenitors across blood endothelium

Project description:T cell recirculation and organ entry involve diapedesis through cytoskeleton remodeling. Part of this process is controlled by guanosine exchange factors (GEFs) and GTPase activating proteins (GAPs) that promote active and inactive forms of Rho family of GTPases. Among GAPs, we identified the ARHGAP45, also known as HMHA1 (human minor histocompatibility antigen 1) and determined its interactome by affinity purification coupled to quantitative mass spectrometry (AP-MS) in Jurkat cells. This dataset contains results of AP-MS of Jurkat cells expressing One-Strep-tagged (OST) ARHGAP45 prior to and after TCR stimulation. Each AP-MS purification of OST- protein is associated with a corresponding control (WT Jurkat) at the same time point of stimulation. For each time point, three independent biological replicates were performed and each biological replicate was analyzed in duplicate by mass spectrometry.

2021-02-04 | PXD018016 | Pride

Identifying the N-terminus of SLX4Nter in HeLa KO30 cells

Project description:The project was aimed at identifying the N-terminus of a truncated form of SLX4 (termed SLX4Nter) in HeLa KO30 cells. The HeLa KO30 cell line comes from a clone of HeLa Flp-In T-Rex cells (termed FITo) obtained through a CRISPR-Cas9 approach using commercially available plasmids from Santa Cruz Biotechnology (sc-404395 and sc-404395-HDR). This allows the integration of an exogenous plasmid conferring Puromycin resistance at the endogenous SLX4 locus. Unexpectedly, this strategy allowed us to retrieve several Puromycin-resistant clones (including KO30) displaying a shorter form of endogenous SLX4 that is N-terminally truncated (SLX4Nter). To identify the proximal peptide of SLX4Nter, FITo and KO30 nuclear extracts were immunoprecipitated with anti-SLX4 antibodies and N-terminus of SLX4Nter was further identified by LC-MSMS analysis.

2022-02-16 | PXD029062 | Pride

RNA-seq of wild type strain and spoIIE deficient Clostridium beijerinckii NCIMB 8052 at 4, 11 and 23 hours of fermentation

Project description:To understand the impact of the stop of the sporulation cycle on the solvent production. We constructed a spoIIE deficient mutant to block the sporulation cycle in the mutant. We performed fermentations and RNA seq analysis to see the impact of the spoIIE disruption on the transcriptome.

2020-04-30 | E-MTAB-7481 | biostudies-arrayexpress

RNA-seq of a degenerated spoIIE deficient Clostridium beijerinckii NCIMB 8052 at 4, 11 and 24 hours of fermentation

Project description:To understand the impact of strain degeneration on the transcriptome we performed fermentations and RNA seq analysis to on a degenerated C.berijeirnckii strain.

2020-06-30 | E-MTAB-9201 | biostudies-arrayexpress

NUPR1 inhibits hyperPARylation-associated cell death

Project description:Nuclear Protein 1 (NUPR1) is a nuclear intrinsically disordered protein of 82 amino acids that plays an important role in stress response and in cancer development. The project was aimed at identifying binding partners of NUPR1 in MiaPaCa-2 cells, in control and after stress condition: glucose starvation during 24h. Lysates from MiaPaCa-2 cells (transfected with NUPR1-Flag or NUPR1-GFP) were used for immunoprecipitation, performed with agarose beads coated with anti-Flag or anti-GFP antibodies. After immunoprecipitation, the eluted proteins were analyzed by LC-MS/MS.

2022-07-29 | PXD035236 | Pride

The dynamic transcriptional profile of sertoli cells during the progression of spermatogenesis

Project description:Sertoli cells (SCs), the only somatic cells within seminiferous tubules, associate intimately with developing germ cells. They not only provide physical and nutritional support but also secrete factors essential to the complex developmental processes of germ cell proliferation and differentiation. The SC transcriptome must therefore adapt rapidly during the different stages of spermatogenesis. We report comprehensive genome-wide expression profiles of pure populations of SCs isolated at 5 distinct stages of the first wave of mouse spermatogenesis, using RNA sequencing technology. We were able to reconstruct about 13 901 high-confidence, nonredundant coding and noncoding transcripts, characterized by complex alternative splicing patterns with more than 45% comprising novel isoforms of known genes. Interestingly, roughly one-fifth (2939) of these genes exhibited a dynamic expression profile reflecting the evolving role of SCs during the progression of spermatogenesis, with stage-specific expression of genes involved in biological processes such as cell cycle regulation, metabolism and energy production, retinoic acid synthesis, and blood-testis barrier biogenesis. Finally, regulatory network analysis identified the transcription factors endothelial PAS domain-containing protein 1 (EPAS1/Hif2α), aryl hydrocarbon receptor nuclear translocator (ARNT/Hif1β), and signal transducer and activator of transcription 1 (STAT1) as potential master regulators driving the SC transcriptional program. Our results highlight the plastic transcriptional landscape of SCs during the progression of spermatogenesis and provide valuable resources to better understand SC function and spermatogenesis and its related disorders, such as male infertility. Genome-wide expression profiling analysis using Illumina next-generation sequencing technology

2015-08-25 | E-GEOD-59698 | biostudies-arrayexpress

Utilization of redundant dBPs in cells with altered levels of MXE regulators

Project description:Usage of intron lariat junctions of human and chicken KGDH transcripts in cells expressing Ca2+-dependent isoforms.

2021-02-07 | E-MTAB-9412 | biostudies-arrayexpress