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Potential ATF6 regulatory proteins were screened by proteomics using pancreatic tissues of PRSS1 transgenic mice treated with caerulein

ABSTRACT: Humanized PRSS1 transgenic mice were constructed and treated with caerulein to mimic the development of human AP and CP. Potential ATF6 regulatory proteins in pancreatitis were screened by proteomics using pancreatic tissues of PRSS1 AP mice.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Pancreas

DISEASE(S): Pancreatitis

SUBMITTER: Zhang guowei

LAB HEAD: Guo-wei Zhang

PROVIDER: PXD016703 | Pride | 2021-09-09

REPOSITORIES: Pride

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	19P02970004_DIA.raw	Raw
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Publications

ATF6 aggravates acinar cell apoptosis and injury by regulating p53/AIFM2 transcription in Severe Acute Pancreatitis.

Tan Jie-Hui JH Cao Rong-Chang RC Zhou Lei L Zhou Zhi-Tao ZT Chen Huo-Ji HJ Xu Jia J Chen Xue-Mei XM Jin Yang-Chen YC Lin Jia-Yu JY Zeng Jun-Ling JL Li Shu-Ji SJ Luo Min M Hu Guo-Dong GD Yang Xiao-Bing XB Jin Jin J Zhang Guo-Wei GW

Theranostics 20200709 18

<b>Background:</b> There is no curative therapy for severe acute pancreatitis (SAP) due to poor understanding of its molecular mechanisms. Endoplasmic reticulum (ER) stress is involved in SAP and increased expression of ATF6 has been detected in SAP patients. Here, we aimed to investigate the role of ATF6 in a preclinical SAP mouse model and characterize its regulatory mechanism. <b>Methods:</b> Pancreatic tissues of healthy and SAP patients were collected during surgery. Humanized PRSS1 transge ...[more]

PMID: 32724472

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Potential regulatory proteins in pancreatitis were screened by proteomics using pancreatic tissues of PRSS1 AP mice

Project description:Humanized PRSS1 transgenic mice were constructed and treated with caerulein to mimic the development of human AP and CP. Potential regulatory proteins in pancreatitis were screened by proteomics using pancreatic tissues of PRSS1 AP mice.

2021-09-09 | PXD016651 | Pride

Quantitative analysis of skin proteomics in Chinese young and old populations by mass spectrometry

Project description:Aging is a normal physiological phenomenon of organisms. Skin aging is a specific manifestation of aging of local human organs. In this study, we used mass spectrometry to perform non-invasive analysis of the skin of 20 healthy young and elderly people in China. Quantitative proteomic analysis identified differentially expressed proteins.

2020-08-05 | PXD018430 | Pride

Unbiased Proteomic Profiling of Mouse Liver Tissue Samples Using HRM™ Mass Spectrometry

Project description:Cryopreserved cGMP-compliant human pluripotent stem cell-derived hepatic progenitors rescue mice from acute liver failure through rapid paracrine effects on liver cells

2024-05-22 | PXD049134 | Pride

Gene expresssion changes in pancreatic islets of 11 weeks old IKK2-CApdx-1 mice compared to control and Pdx-1+/- mice

Project description:Canonical IKK/NF-κB signaling is a master regulator of inflammation and innate immunity and has been implicated in the pathogenesis of T1D. To investigate the impact of NF-κB activation on β-cell homeostasis and diabetes development, we generated a transgenic gain-of-function mouse model allowing conditional NF-κB activation via expression of IKK2-CA (constitutively active IKK2 allele) in β-cells using the tetracycline-regulated gene expression system. Pdx-1-tTA (knockin model generating Pdx-1 haploinsufficiency) driver mice were used for β-cell specific transgene expression. Double transgenic IKK2-CA-pdx-1 mice develop a full-blown immune-mediated diabetes.To identify gene expression changes underlying this diabetes development pancreatic islets of diabetic IKK2-CA-Pdx-1, PDX-1 +/- and control mice were prepared and isolated total RNA was used for microarray analysis. Pancreatic islets were isolated from 11 weeks old control (n=5), Pdx-1+/- (n=4) and IKK2-CApdx-1 (n=5) mice, total RNA was extracted with RNeasy mini kit (QIAGEN). Microarray analyses were performed using 200 ng total RNA as starting material and 5.5 µg ssDNA per hybridization (GeneChip Fluidics Station 450; Affymetrix, Santa Clara, CA). The total RNAs were amplified and labeled following the Whole Transcript (WT) Sense Target Labeling Assay (http://www.affymetrix.com). Labeled ssDNA was hybridized to Mouse Gene 1.0 ST Affymetrix GeneChip arrays (Affymetrix, Santa Clara, CA). The chips were scanned with a Affymetrix GeneChip Scanner 3000 and subsequent images analyzed using Affymetrix® Expression Console Software (Affymetrix). Probe level data were obtained using the robust multichip average (RMA) normalization algorithm.

2014-05-02 | E-GEOD-47504 | biostudies-arrayexpress

APPLICATION OF A NOVEL PEPTIDE SPECTRAL LIBRARY USING SWATH ANALYSIS FOR THE QUANTITATION OF PROTEINS IN EX-DIAGNOSTIC CLINICAL SERUM SAMPLES FROM SHEEP

Project description:This project applies SWATH approach to analyse ex-diagnostic sheep serum samples from actual clinical cases submitted to The University of Queensland's School of Veterinary Science, Veterinary Laboratory Services (VLS) compared with serum of normal sheep.

2022-11-15 | PXD005077 | Pride

Endogenous pull-downs of human proteins ATP6V1A and AHNAK to validate interactions with influenza A virus proteins M2 and NEP by targeted mass spectrometry in virus-infected cells

Project description:Influenza A Virus (IAV) is a recurring respiratory virus with antiviral therapies of limited use. Understanding host proteins essential for IAV infection can identify targets for alternative host-directed therapies (HDTs). Using affinity purification-mass spectrometry (AP-MS) and global phosphoproteomic and protein abundance analyses with three IAV strains (pH1N1, H3N2, H5N1) in three human cell types (A549, NHBE, THP-1), we mapped 332 IAV-human protein-protein interactions (PPIs) and identified 13 IAV-modulated kinases. Whole exome sequencing of patients who experienced severe influenza revealed several genes, including the structural scaffold protein AHNAK, with predicted loss-of-function variants that were also identified in our proteomic analyses. Of our identified host factors, 54 significantly altered IAV infection upon siRNA knockdown, and two factors, COPB1 and AHNAK, were also essential for productive infection by SARS-CoV-2. Finally, 16 compounds targeting our identified host factors suppressed IAV replication, with two targeting CDK2 and FLT3 showing pan-antiviral activity across influenza and coronavirus families. This study provides a comprehensive network model of IAV infection in human cells, identifying functional host targets for pan-viral HDT. This project includes endogenous IP data from A549 cells and NHBE cells infected with H5N1 IAV validating IAV-host PPIs M2-ATP6V1A and NEP-AHNAK, respectively. M2-ATP6V1A and NEP-AHNAK PPIs were first identified in AP-MS data that is submitted under its own dataset identifier (PXD036077).

2023-07-25 | PXD041663 | Pride

Mapping the protein interaction network of adaptor protein complex in Arabidopsis identifies P34 as a common stability regulator

Project description:Adaptor protein (AP) complexes are evolutionarily conserved vesicle transport regulators that recruit coat proteins, membrane cargos and coated vesicle accessory proteins. Since in plants endocytic and post-Golgi trafficking intersect at the trans-Golgi network, unique mechanisms for sorting cargos of overlapping vesicular routes are anticipated. The plant AP complexes are part of the sorting machinery, but despite some functional information, their cargoes, accessory proteins, and regulation remain largely unknown. Here, by means of various proteomics approaches, we generated the overall interactome of the five AP and the TPLATE complexes in Arabidopsis thaliana. The interactome converged on a number of hub proteins, including the thus far unknown adaptin binding-like protein, designated P34. P34 interacted with the clathrin-associated AP complexes, controlled their stability and, subsequently, influenced clathrin-mediated endocytosis and various post-Golgi trafficking routes. Altogether, the AP interactome network offers substantial resources for further discoveries of unknown endomembrane trafficking regulators in plant cells.

2023-12-07 | PXD035454 | Pride

Analysis of context-specific KRAS-effectors (sub)complexes in Caco-2 cells

Project description:Ras is a key switch controlling cell behavior. In the GTP-bound form, Ras interacts with numerous effectors in a mutually exclusive manner, where individual Ras-effectors are likely part of larger cellular (sub)complexes. The molecular details of these (sub)complexes and their alteration in specific contexts is not understood. Focusing on KRAS, we performed affinity purification (AP)-mass spectrometry (MS) experiments of exogenous expressed Flag-KRAS WT and three oncogenic mutants (‘genetic contexts’) in the human Caco-2 cell line, each exposed to 11 different culture media (‘culture contexts’) that mimic conditions relevant in the colon and colorectal cancer. We identified four effectors present in complex with KRAS in all genetic and growth contexts (‘context-general effectors’). Seven effectors are found in KRAS complexes in only some contexts (‘context-specific effectors’). Analyzing all interactors in complex with KRAS per condition, we find that the culture contexts had a larger impact on interaction rewiring than genetic contexts. We analyzed how changes in the interactome impact functional outcomes and created interactive shiny app for interactive visualization. We validated some of the functional differences in metabolism and proliferation. Finally, we used networks to evaluate how KRAS effectors are involved in the modulation of functions by random walk analyses of effector-mediated (sub)complexes. Altogether, our work shows the impact of environmental contexts on network rewiring, which provides insides into tissue-specific signaling mechanisms. This may also explain why KRAS oncogenic mutants may be causing cancer only in specific tissues despite KRAS being expressed in most cells and tissues.

2023-02-23 | PXD035399 | Pride

Oncogenic ETS proteins regulate a Ras/MAPK gene expression program in the absence of MAPK signaling

Project description:Approximately 50% of prostate cancers have chromosomal translocations resulting in the over-expression one of four ETS family transcription factors. However, it is not known why these four four family members are selected for oncogenic roles, while other ETS proteins are not. We found that the four oncogenic ETS family members have a specific role in prostate cell migration. Using chromatin immunoprecipitation coupled with next-generation sequencing, this specific biological function was matched to a specific set of genomic targets highlighted by the presence of an AP-1 binding site. ETS/AP-1 binding sites are prototypical Ras-responsive elements, but oncogenic ETS proteins could activate a Ras/MAPK transcriptional program in the absence of MAPK activation. These findings indicate that the specific function of ETS proteins over-expressed in prostate cancer is the activation of a Ras/MAPK gene expression program in the absence of signaling pathway mutations. 16 samples were analyzed, comprised of four replicates each of four different biological conditions. RNA from U0126 treated RWPE-1 empty vector cell RNA serves as a control for each experiment. Cell lines have retroviral expression constructs expressing either empty vector, Flag-ERG, or Flag-ETV1.

2011-10-18 | E-GEOD-29438 | biostudies-arrayexpress

Analalysis of ASPP2 interaction

Project description:Quantitative AP-MS from HEK 293T cells expressing Strep-SHA tagged ASPP2 or ASPP2. (Ex1) Quantitative AP-MS from HEK293 cells expressing Strep-SHA tagged PP1alpha, beta, gamma. (Ex2)

2019-04-02 | PXD012378 | Pride

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