Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

The Whole cell lysate and Crude membrane of HEK293T cells

ABSTRACT: We perform the experiment in order to compare identified membrane proteins from whole cell lysate and membrane fraction in HEK293T cell. The isolation of membrane fractions is critical to obtain a more comprehensive membrane proteome, especially with proteins containing transmembrane domains.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Shushi Nagamori

PROVIDER: PXD027472 | JPOST Repository | Thu Apr 28 00:00:00 BST 2022

REPOSITORIES: jPOST

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	1.1_MF_f0.raw	Raw
	1.1_MF_f1.raw	Raw
	1.1_MF_f2.raw	Raw
	1.1_MF_f3.raw	Raw
	1.2_MF_f0.raw	Raw

Items per page:

1 - 5 of 18

Publications

Simple But Efficacious Enrichment of Integral Membrane Proteins and Their Interactions for In-Depth Membrane Proteomics.

Kongpracha Pornparn P Wiriyasermkul Pattama P Isozumi Noriyoshi N Moriyama Satomi S Kanai Yoshikatsu Y Nagamori Shushi S

Molecular & cellular proteomics : MCP 20220125 5

Membrane proteins play essential roles in various cellular processes, such as nutrient transport, bioenergetic processes, cell adhesion, and signal transduction. Proteomics is one of the key approaches to exploring membrane proteins comprehensively. Bottom-up proteomics using LC-MS/MS has been widely used in membrane proteomics. However, the low abundance and hydrophobic features of membrane proteins, especially integral membrane proteins, make it difficult to handle the proteins and are the bot ...[more]

PMID: 35085786

Similar Datasets

The Whole cell lysate and Crude membrane of HEK293T cells

Project description:We perform the experiment in order to compare identified membrane proteins from whole cell lysate and membrane fraction in HEK293T cell. The isolation of membrane fractions is critical to obtain a more comprehensive membrane proteome, especially with proteins containing transmembrane domains.

2024-09-10 | PXD027459 | JPOST Repository

Whole-genome landscape PARP1 in HEK293T cells

Project description:Poly [ADP-ribose] polymerase 1 (PARP1) is involved in differentiation, proliferation, tumor transformation, in repair of single-stranded DNA and double-stranded DNA breaks (DSBs) in conjunction with BRCA. PARP1 enzyme works modifying nuclear proteins by poly ADP-ribosylation. It was found that PARP1 and HNRNPA2B1 bind at termini of forum domains and may play a role in coordinated regulation of genes in forum domains (Tchurikov et al., 2013). Forum domains are long DNA fragments of excised chromosomal DNA. Mostly forum domains are of 50-200 kb in length, although larger domains, up to 500 - 700 kb, are also observed. The domains are delimited by hot spots of double-strand breaks (DSBs). This ChIP-Seq is aimed to compare genome-wide binding sites of PARP1 with different genomic features, including the hot spots of DSBs.

2017-02-01 | GSE74954 | GEO

Membrane-associated transcripts in Arabidopsis; their isolation...

Project description:Membrane-associated, integral membrane and secreted proteins are of key importance in many cellular processes. For most of the 28 952 predicted proteins in Arabidopsis, the actual subcellular localisation has not been demonstrated experimentally. So far, their potential membrane-association has been deduced from algorithms that predict transmembrane domains and signal peptides. However, the comprehensiveness and accuracy of these algorithms is still limited. The majority of membrane-associated and secreted proteins is synthesised on membrane-bound polysomes. Therefore, the isolation and characterisation of mRNA associated with membrane-bound polysomes offers an experimental tool for the genome-wide identification of these proteins. Here we describe an efficient method to isolate mRNA from membrane-bound polysomes and report on the validation of the method to enrich for transcripts encoding membrane-associated and secreted proteins. The sensitivity and reproducibility of the isolation method was investigated by DNA microarray analysis. Pearson correlations between transcript levels obtained from three replicate isolations showed that the method is highly reproducible. A significant enrichment for mRNAs encoding proteins containing predicted transmembrane domains and signal peptides was observed in the membrane-bound polysomal fraction. In this fraction, 301 transcripts were classified by gene ontologies as âcellular component unknownâ, and potentially encode previously unrecognised secreted or membrane-associated proteins. Membrane-bound (MBP) and free polysomes (FP) were isolated in triplicate from one batch of two weeks old aboveground parts of Arabidopsis seedlings. Each combination of isolated MBP and FP was applied to 4 microarrays, of which two were dyeswapped, giving a total of 12 arrays.

2010-07-01 | E-GEOD-4023 | biostudies-arrayexpress

Next-generation RNA sequencing of human astrocyte cultures to study the comparative levels of transmembrane proteins and their transcriptional variants

Project description:The aim of this study was to perform Next-generation RNA sequencing to analyse the expression of transmembrane proteins in human astrocytes and compare the prevalance of transcriptional variants thereof.

2023-05-25 | GSE228084 | GEO

ChIP-seq experiment in J20 hAPP mice and Pilocarpine-treated mice using deltaFosB antibody against whole-hippocampal lysate

Project description:∆FosB epigenetically regulates target gene expression; however, whether ∆FosB targets the same genes when induced by recurrent seizures in different neurological conditions like Alzheimer's disease (AD) is unclear. We performed hippocampal ChIP-sequencing to identify ∆FosB target genes in APP mice, a model of AD, and in pilocarpine-treated wildtype mice (Pilo mice), a model of epilepsy. Functional domains modulated by ∆FosB target genes are expanded and diversified in APP mice and in Pilo mice (vs respective controls), and primarily involve neuronal excitability and neurotransmission, neurogenesis, chromatin remodeling, and cellular stress and immunity. To identify genes bound by ∆FosB regardless of seizure etiology, we focused on the 442 ∆FosB target genes in both APP and Pilo mice (not respective controls), which related to pathways including membrane potential regulation, glutamatergic signaling, complement activation, neuron-glia population maintenance, and chromatin dynamics. RNA-sequencing and RT-qPCR in independent mice detected altered expression of several ∆FosB targets shared in APP and Pilo mice. Our findings indicate that seizure-induced ∆FosB can bind genes in seizure etiology-dependent patterns, but can also bind genes in patterns regardless of seizure etiology. Understanding factors that influence ∆FosB binding patterns could reveal novel insights into control of gene expression in disorders with recurrent seizures.

2020-04-16 | E-MTAB-8954 | biostudies-arrayexpress

Membrane-associated transcripts in Arabidopsis; their isolation...

Project description:Membrane-associated, integral membrane and secreted proteins are of key importance in many cellular processes. For most of the 28 952 predicted proteins in Arabidopsis, the actual subcellular localisation has not been demonstrated experimentally. So far, their potential membrane-association has been deduced from algorithms that predict transmembrane domains and signal peptides. However, the comprehensiveness and accuracy of these algorithms is still limited. The majority of membrane-associated and secreted proteins is synthesised on membrane-bound polysomes. Therefore, the isolation and characterisation of mRNA associated with membrane-bound polysomes offers an experimental tool for the genome-wide identification of these proteins. Here we describe an efficient method to isolate mRNA from membrane-bound polysomes and report on the validation of the method to enrich for transcripts encoding membrane-associated and secreted proteins. The sensitivity and reproducibility of the isolation method was investigated by DNA microarray analysis. Pearson correlations between transcript levels obtained from three replicate isolations showed that the method is highly reproducible. A significant enrichment for mRNAs encoding proteins containing predicted transmembrane domains and signal peptides was observed in the membrane-bound polysomal fraction. In this fraction, 301 transcripts were classified by gene ontologies as ‘cellular component unknown’, and potentially encode previously unrecognised secreted or membrane-associated proteins. Keywords: mRNA localisation, mRNA fractionation, predict protein localisation

2006-03-17 | GSE4023 | GEO

The ER membrane protein complex interacts cotranslationally to enable biogenesis of multipass membrane proteins

Project description:The endoplasmic reticulum (ER) supports biosynthesis of proteins with diverse transmembrane domain (TMD) lengths and hydrophobicity. Features in transmembrane domains such as charged residues in ion channels are often functionally important, but could pose a challenge during cotranslational membrane insertion and folding. Our systematic proteomic approaches in both yeast and human cells revealed that the ER membrane protein complex (EMC) binds to and promotes the biogenesis of a range of multipass transmembrane proteins, with a particular enrichment for transporters. Proximity-specific ribosome profiling demonstrates that the EMC engages clients cotranslationally and immediately following clusters of TMDs enriched for charged residues. The EMC can remain associated after completion of translation, which both protects clients from premature proteasomal degradation and allows recruitment of substrate-specific and general chaperones. Thus, the EMC broadly enables the biogenesis of multipass transmembrane proteins containing destabilizing features, thereby mitigating the trade-off between function and stability.

2018-05-29 | GSE112891 | GEO

Transcriptomic responses to SEL1L-HRD1 ERAD deficiency in HEK293T cells and mouse brown adipose tissue

Project description:The mamallian SEL1L-HRD1 endoplasmic reticulum-associated degradation (ERAD) complex is essential for retrotranslocation and degradatin of misfolded proteins in the ER. HRD1 is an E3-ligase with multiple transmembrane domains that consititute the retrotranslocation channel, while its cofactor SEL1L is responsible for the stability of the complex and for substrate recruitment. This study aims to identify transcriptionally-regulated genes responding to SEL1L-HRD1 ERAD deficiency in two different mammalian cell types.

2023-12-17 | GSE231583 | GEO

Top-down proteomics of membrane proteoforms from HeLa cells and mouse hippocampus

Project description:Comprehensive characterization of membrane proteins at the level of proteoforms in complex biological samples by top-down mass spectrometry is of vital importance in revealing their precise functions. Based on a novel membrane protein separation technology, we obtained a dataset of membrane proteoforms from HeLa cells and mouse hippocampus, containing information about combinatorial post-translational modifications, truncation, and complete transmembrane domains, which accelerate our understanding and exploitation of membrane proteins.

2023-02-08 | PXD039949 |

N-Linked Glycoprotein Profiles of Human Breast Basal Malignant Cell Lines

Project description:Secreted proteins and transmembrane proteins with extracellular domains are frequently glycosylated; this group of proteins includes those that participate in the various intercellular junctions and signaling pathways of an epithelium. In this study we characterized the differences in glycoprotein expression between claudin-low and other breast cell lines using a dataset of 26 breast cell lines in which the glycoproteins were identified and quantitated by liquid chromatography/ tandem mass spectrometry. Our goals are to characterize the glycoproteome of a set of claudin-low lines, compare them to basal, luminal and non-malignant cells and to identify drugs that may be especially effective on these cell lines. These nine basal malignant breast cells (HCC1143, HCC1395, HCC1937, HCC1954, HCC38, HCC70, MDAMB468, SUM149PT and SUM229PE) data are a part of 26 breast cell lines we analyzed.

2017-03-27 | PXD005295 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data