ABSTRACT: Untargeted LC-TIMS-MS/MS dataset collected on Bruker timsTOF fleX for Penicillium atramentosum RS17, meleagrin standard, and roquefortine C standard.
Project description:Untargeted LC-TIMS-MS/MS dataset collected on Bruker timsTOF fleX for Penicillium atramentosum RS17, meleagrin standard, and roquefortine C standard.
Project description:The Enhancer of Zeste 2 Polycomb Repressive Complex 2 Subunit (EZH2) is an essential epigenetic modifier able to methylate lysine 27 on histone H3 (H3K27) to induce chromatin compaction, protein complex recruitment and ultimately transcriptional repression. Hematologic malignancies, including Diffuse Large B cell lymphoma (DLBCL) and Acute myeloid leukemia (AML) have shown a high EZH2-mutation frequency (>20%) associated with poor clinical outcomes. Particularly, two distinct oncogenic mutations, so-called gain-of-function (Y641F and A677G) and loss-of-function (H689A and F667I) are found in the catalytic domain of EZH2. In this study, a comprehensive multi-omics approach was employed to characterize downstream effects of H3K27me3 deposition driven by EZH2 mutations. Human embryonic kidney cells (HEK293T) were transfected to generate three stable EZH2 mutants: EZH2(Y641F), EZH2(A677G), and EZH2(H689A/F667I), which were validated via immunoblotting and DIA-MS-based histone profiling assay. Subsequently, constructs were analyzed under a comprehensive multi-omics approach including: 1) Cleavage Under Targets and Tagmentation (CUT&Tag); 2) chromatin accessibility characterization using the assay for transposase-accessible chromatin with sequencing (ATAC-Seq); 3) transcriptomics (RNA-Seq); 4) label-free whole-cell proteomics, acquired with a Bruker timsTOF Pro HPLC-MS/MS with Ion Mobility, and 5) MS-based untargeted metabolomics, in positive and negative ionization MS/MS mode, acquired with an Agilent 6545 QTOF with a 1290 UHPLC system and HILIC column. Total coverage comprised over 21,000 chromatin regions, 18,000 transcripts, 6,000 proteins and 400 metabolic features. Pair-wise comparison using univariate and supervised multivariate statistical methods revealed significant differences between constructs in each omic level. Furthermore, effector pathway analysis combining omics data revealed distinctive reprogramming effects for each EZH2 mutant.
Project description:The Enhancer of Zeste 2 Polycomb Repressive Complex 2 Subunit (EZH2) is an essential epigenetic modifier able to methylate lysine 27 on histone H3 (H3K27) to induce chromatin compaction, protein complex recruitment and ultimately transcriptional repression. Hematologic malignancies, including Diffuse Large B cell lymphoma (DLBCL) and Acute myeloid leukemia (AML) have shown a high EZH2-mutation frequency (>20%) associated with poor clinical outcomes. Particularly, two distinct oncogenic mutations, so-called gain-of-function (Y641F and A677G) and loss-of-function (H689A and F667I) are found in the catalytic domain of EZH2. In this study, a comprehensive multi-omics approach was employed to characterize downstream effects of H3K27me3 deposition driven by EZH2 mutations. Human embryonic kidney cells (HEK293T) were transfected to generate three stable EZH2 mutants: EZH2(Y641F), EZH2(A677G), and EZH2(H689A/F667I), which were validated via immunoblotting and DIA-MS-based histone profiling assay. Subsequently, constructs were analyzed under a comprehensive multi-omics approach including: 1) Cleavage Under Targets and Tagmentation (CUT&Tag); 2) chromatin accessibility characterization using the assay for transposase-accessible chromatin with sequencing (ATAC-Seq); 3) transcriptomics (RNA-Seq); 4) label-free whole-cell proteomics, acquired with a Bruker timsTOF Pro HPLC-MS/MS with Ion Mobility, and 5) MS-based untargeted metabolomics, in positive and negative ionization MS/MS mode, acquired with an Agilent 6545 QTOF with a 1290 UHPLC system and HILIC column. Total coverage comprised over 21,000 chromatin regions, 18,000 transcripts, 6,000 proteins and 400 metabolic features. Pair-wise comparison using univariate and supervised multivariate statistical methods revealed significant differences between constructs in each omic level. Furthermore, effector pathway analysis combining omics data revealed distinctive reprogramming effects for each EZH2 mutant.
Project description:The Enhancer of Zeste 2 Polycomb Repressive Complex 2 Subunit (EZH2) is an essential epigenetic modifier able to methylate lysine 27 on histone H3 (H3K27) to induce chromatin compaction, protein complex recruitment and ultimately transcriptional repression. Hematologic malignancies, including Diffuse Large B cell lymphoma (DLBCL) and Acute myeloid leukemia (AML) have shown a high EZH2-mutation frequency (>20%) associated with poor clinical outcomes. Particularly, two distinct oncogenic mutations, so-called gain-of-function (Y641F and A677G) and loss-of-function (H689A and F667I) are found in the catalytic domain of EZH2. In this study, a comprehensive multi-omics approach was employed to characterize downstream effects of H3K27me3 deposition driven by EZH2 mutations. Human embryonic kidney cells (HEK293T) were transfected to generate three stable EZH2 mutants: EZH2(Y641F), EZH2(A677G), and EZH2(H689A/F667I), which were validated via immunoblotting and DIA-MS-based histone profiling assay. Subsequently, constructs were analyzed under a comprehensive multi-omics approach including: 1) Cleavage Under Targets and Tagmentation (CUT&Tag); 2) chromatin accessibility characterization using the assay for transposase-accessible chromatin with sequencing (ATAC-Seq); 3) transcriptomics (RNA-Seq); 4) label-free whole-cell proteomics, acquired with a Bruker timsTOF Pro HPLC-MS/MS with Ion Mobility, and 5) MS-based untargeted metabolomics, in positive and negative ionization MS/MS mode, acquired with an Agilent 6545 QTOF with a 1290 UHPLC system and HILIC column. Total coverage comprised over 21,000 chromatin regions, 18,000 transcripts, 6,000 proteins and 400 metabolic features. Pair-wise comparison using univariate and supervised multivariate statistical methods revealed significant differences between constructs in each omic level. Furthermore, effector pathway analysis combining omics data revealed distinctive reprogramming effects for each EZH2 mutant.
Project description:LC-MS(/MS) and MALDI-(TIMS)-qTOF imaging MS(/MS) datasets for the fungal-fungal interaction between Penicillium sp. JBC and Diutina catenulata 135E/Brevibacterium linens JB5. MALDI-TIMS-qTOF MS/MS data for a purchased standard of sclerotigenin is included.
Project description:Small molecule profiles for cheese rind microbiome isolates collected for molecular association network analysis via IDBac. Small molecule profiles for four biological replicates were collected for each strain grown on 10% cheese curd agar on a Bruker timsTOF fleX after 7 days of growth at room temperature.
Project description:A comprehensive LFQ benchmark dataset to validate data analysis pipelines on modern day acquisition strategies in proteomics using SCIEX TripleTOF5600 and 6600+, Orbitrap QE-HFX, Waters Synapt GS-Si and Synapt XS and Bruker timsTOF Pro.
Project description:The present study was initiated to evaluate the quantitative proteomic profiling of Protothecazopfiigenotypes. The cells (P.zopfii genotype 1 –noninfectious and genotype 2 -infectious) were cultured in triplicates until the mid-logarithmic growth phase; the proteins were extracted after sonication on ice, followed by 2D DIGE separation as recommended by the manufacturer (GE Healthcare). The differentially expressed proteins spots were identified using Decodon software analysis (Delta 2D version 4.0 software). Protein identification was carried out by MALDI TOF MS/MS (Ultraflex III TOF/TOF, Bruker Daltonics, Bremen, Germany). The spectra was acquired using the automated option (AutoXecute ) of the Flex Analysis software version 3.3 (Bruker Daltonics, Leipzig, Germany), processed using Flex analysis software version 3.3 (Bruker Daltonics, Leipzig, Germany) and the database search was conducted using the MS/MS ion search (MASCOT, http://www.matrixscience.com) against all entries of NCBInr (GenBank)/Swissprot with subsequent parameters: trypsin digestion, up to one missed cleavage, fixed modifications: carbamidomethyl and with the following variable modifications: oxidation (M), peptide tol.: +- 1.2 Da, MS/MS tol.: +- 0.6 Da, peptide charge: +1. The results were assessed using MOWSE score, p- and E values and those possess positive hits with cRAP database were eliminated from the list.
Project description:Human follicular fluid samples were spiked with 50uM norepinephrine-D6. The 'spiked' sample was also spiked with 50uM norepinephrine. Samples were washed using a BondElut PBA column and analyzed using LC-MS on a Bruker timsTOF fleX.
Project description:Single H358 cells analyzed using SCOPE2 on a TIMSTOF Flex mass spectrometer. Bruker .d folders, MGFs, Proteome Discoverer 2.5 and MaxQuant 1.6.17 results are uploaded.