Project description:Comparative interactome analysis of Nematostella vectensis Hsp70 isoforms reveals unique clientome remodeling upon heat stress. Three isoforms of nvHSP70 (A, B, and D) were IP'd from either Heat Shock conditions (HS) or non-Heat Shock (UN) and their interactomes were compared. Mass Spec was run on a Q-Exactive (Orbitrap).
Project description:The project aimed to characterize the protein composition from 3 Pleistocene bone specimens (AR-7, AR-16, AR-30). Analysed extracts concern standard ZooMS tryptic digests (AR-7, AR-16, AR-30) and additional palaeoproteomic extracts (AR-30A and AR-30B). The latter concern two biological duplicates. All extractions were performed at the Department of Human Evolution, MPI-EVA (Germany) under sterile conditions. Analyses took place on a Q-Exactive Hybrid Quadrupole-Orbitrap MS.
Project description:Proteomics and transcriptomics data of tomato fruits (Solanum lycopersicum L. var. Moneymaker) at 9 developmental stages were used to calculate with a mathematical model the rate constants of synthesis and degradation for over 1,000 proteins. Proteome and transcriptome were extracted from the pericarp tissue and analyzed using label-free LC-MS/MS (Orbitrap Q-Exactive) and RNA Sequencing (Illumina), respectively. Absolute quantification of transcriptome has been obtained by spiking-in internal standard before total-RNA extraction. Absolute quantification of the proteome has been approximated using the "Total Protein" approach. An OD equation defining the changes of protein content has been used to determine the synthesis and degradation rate constants (day -1). Almost 2,400 transcript-protein pairs were identified and the translation and degradation rate constants were determined for more than a thousand proteins. The model predicted median values of about 2 min for the translation and a lifetime of approximately 11 days. Proteins involved in protein synthesis had higher ks and kd values, indicating that the protein machinery is particularly flexible. None sequenced-based features were found that could be used to predict these rate constants.
Project description:Proteomics and phosphoproteomics analysis of MCF7 cells sensitive and resistant to the PI3K inhibitor GDC-0941. The analysis was done for parental and 3 resistant cell lines maintained in the presence or the absence of the drug. One of the resistant cell lines (G2) was also treated with vehicle or the kinase inhibitors GDC0941, MK-2206 and Ku-0063794 for 2h. The proteomics analysis of sensitive and resistant MCF7 cells in basal conditions and phosphoproteomics analysis of the G2 cells in the presence of inhibitors of the PI3K pathway were run in a nano flow ultrahigh pressure liquid chromatography (UPLC, nano Acquity, Waters) coupled to an LTQ-Orbitrap XL mass spectrometer (Thermo Fisher Scientific). The phosphoproteomics study of sensitive and resistant MCF7 cells in basal conditions was run in a Dionex UltiMate 3000 RSLCnano coupled to an Orbitrap Q Exactive Plus mass spectrometer (Thermo Fisher Scientific).
Project description:To explore the cellular pathways and the molecular functions affected by a novel biogenic amine, (3-HKA), we performed a label free quantitative (LFQ) proteomic analysis of mouse lymphatic endothelial cells (LEC), and primary dendritic cells, treated with IFNgammaplus or minus 3-HKA. Biological triplicates, of total cell lysates, were fractionated by one-dimensional gel electrophoresis (1DEF) and the “in gel” tryptic derived peptides analyzed by nano-LC-ESI-MS/MS on a Q Exactive HF quadrupole orbitrap mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). LFQ analysis highlighted that 3-HKA downregulated many of the inflammatory pathways, more notably JAK/STAT1 and NFkB, associated with IFNgamma activation.
Project description:Macrophages were exposed to CFNA from different blood products prior to gene expression array: Red Blood Cells units (RBC), Fresh Frozen Plasma Units (FFP) and Platelet Concentrates (PC). Extracts from PBS were used as controls. Biological triplicates were included with three different macrophage cell lines (M1, M2, M3).
Project description:This study investigated the consumption of milk products in the archaeological record, utilizing human dental calculus as a reservoir of dietary proteins from archaeological samples from across Eurasia. Protein extraction and generation of tryptic peptides from dental calculus was performed using a filter-aided sample preparation (FASP) protocol, modified for ancient samples, on 92 samples of archaeological dental calculus. Samples were extracted at three laboratories; the Functional Genomics Centre Zürich (FGCZ), the Centre for GeoGenetics at the National History Museum of Denmark, and BioArCh at the University of York. Sample extracts were sequenced (LC-MS/MS) using an LTQ-Orbitrap Velos (FGCZ), a Q-Exactive Hybrid Quadrupole Orbitrap and an LTQ-Orbitrap Elite (Central Proteomics Facility, Target Discovery Institute, Oxford).
Project description:This study investigated the consumption of milk products in the archaeological record, utilizing human dental calculus as a reservoir of dietary proteins from archaeological samples from across Eurasia. Protein extraction and generation of tryptic peptides from dental calculus was performed using a filter-aided sample preparation (FASP) protocol, modified for ancient samples, on 92 samples of archaeological dental calculus. Samples were extracted at three laboratories; the Functional Genomics Centre Zürich (FGCZ), the Centre for GeoGenetics at the National History Museum of Denmark, and BioArCh at the University of York. Sample extracts were sequenced (LC-MS/MS) using an LTQ-Orbitrap Velos (FGCZ), a Q-Exactive Hybrid Quadrupole Orbitrap and an LTQ-Orbitrap Elite (Central Proteomics Facility, Target Discovery Institute, Oxford).
Project description:This study investigated the consumption of milk products in the archaeological record, utilizing human dental calculus as a reservoir of dietary proteins from archaeological samples from across Eurasia. Protein extraction and generation of tryptic peptides from dental calculus was performed using a filter-aided sample preparation (FASP) protocol, modified for ancient samples, on 92 samples of archaeological dental calculus. Samples were extracted at three laboratories; the Functional Genomics Centre Zürich (FGCZ), the Centre for GeoGenetics at the National History Museum of Denmark, and BioArCh at the University of York. Sample extracts were sequenced (LC-MS/MS) using an LTQ-Orbitrap Velos (FGCZ), a Q-Exactive Hybrid Quadrupole Orbitrap and an LTQ-Orbitrap Elite (Central Proteomics Facility, Target Discovery Institute, Oxford).
Project description:This study investigated the consumption of milk products in the archaeological record, utilizing human dental calculus as a reservoir of dietary proteins from archaeological samples from across Eurasia. Protein extraction and generation of tryptic peptides from dental calculus was performed using a filter-aided sample preparation (FASP) protocol, modified for ancient samples, on 92 samples of archaeological dental calculus. Samples were extracted at three laboratories; the Functional Genomics Centre Zürich (FGCZ), the Centre for GeoGenetics at the National History Museum of Denmark, and BioArCh at the University of York. Sample extracts were sequenced (LC-MS/MS) using an LTQ-Orbitrap Velos (FGCZ), a Q-Exactive Hybrid Quadrupole Orbitrap and an LTQ-Orbitrap Elite (Central Proteomics Facility, Target Discovery Institute, Oxford).