Project description:UVPD was implemented on an Orbitrap Q-Exactive plus equipped with a ESI/EP-MALDI. UVPD of MALDI generated ions was benchmarked against MALDI-ISD, MALDI-HCD and ESI-UVPD. MALDI UVPD outperformed MALDI HCD and ISD efficiently sequencing small proteins ions.
Project description:The project aimed to identify interaction sites of RNF168 with the Nucleosomes. RNF168 interaction with the Nucleosome was probed by BS3 crosslinking to support structure modeling based on NMR and mutagenesis experiments
Project description:In this study we compared three different fragmentation techniques and two combined fragmentation schemes available on a novel tribrid mass spectrometer (Orbitrap Fusion Lumos, Themro Fisher Scientific) CID, HCD, ETD, ETD with supplemental CID (ETciD) and ETD with supplemental HCD (EThcD) on cross-linked peptides obtained by tryptic cleavage of SDA-cross-linked Human Serum Albumin (HSA). The three-dimensional structure of HSA has been resolved by X-ray crystallography [35] and is used as ground-truth to evaluate the identification results. Right choice of the fragmentation method allows increasing the number of identified linkage sites, increasing the sequence coverage of both linked peptides thereby reducing the second peptide problem, and increasing the precision of cross-link site calling.
Project description:BS3 Crosslinking experiments were performed on in vitro preparations of histone methyltransferase dimers. Each sample was fractionated into two size-exclusion chromatography fractions that were acquired by alternating HCD/EThcD scans of the same precursor on a Fusion-Lumos instrument.
File mappings:
GLP homodimer, SEC1: Z20210712-08
GLP homodimer, SEC2: Z20210712-09
G9a homodimer, SEC1: Z20210712-10
GLP homodimer, SEC2: Z20210712-11
G9aGLP heterodimer, SEC1: Z20210712-12
G9aGLP heterodimer, SEC2: Z20210712-13
Project description:The data sets of human IgG and human fibrinogen are used as test data for the publication of glyXtoolMS (https://github.com/glyXera/glyXtoolMS). GlyXtoolMS is an open-source analysis software for the (semi)automated targeted analysis of glycopeptide mass spectrometry data using OpenMS/TOPPAS as a framework and pipeline engine. The proteins were selected to show the successful analysis of both N-glycopeptide and O-glycopeptide samples. The samples were measured by nano reversed phase liquid chromatography coupled online to an electrospray ionization orbitrap mass spectrometer (nano RP-LC ESI- OT-MS/MS; LTQ Orbitrap Elite, Thermo Scientific, Waltham, MA, USA) with HCD fragmentation.
Project description:The Nucleosome Remodeling and Deacetylase (NuRD) complex is essential for development in complex animals but has been refractory to detailed analysis because of its low abundance and resistance to recombinant production. In combination with other techniques, crosslinking mass spectrometry was used to elucidate the structure of the Nucleosome Remodelling and Deacetylase (NuRD) complex. Natively-purified NuRD, and four recombinantly-expressed NuRD subcomplexes, and a PWWP2A-MTA-HDAC-RBBP alternative ‘NuRD-like’ complex were subjected to crosslinking with DSS, ADH, BS3 and DMTMM.