A5074927619- Advanced Proteomics Techniques and Applications
- Mass Spectrometry Techniques and Applications
- T-cell and B-cell Immunology
- Metabolomics and Mass Spectrometry Studies
- Immune Cell Function and Interaction
- Escherichia coli research studies
- Immunotherapy and Immune Responses
- vaccines and immunoinformatics approaches
- Monoclonal and Polyclonal Antibodies Research
- Glycosylation and Glycoproteins Research
- Virus-based gene therapy research
- Cervical Cancer and HPV Research
- Ubiquitin and proteasome pathways
- Advanced biosensing and bioanalysis techniques
- Photosynthetic Processes and Mechanisms
- Cytokine Signaling Pathways and Interactions
- Receptor Mechanisms and Signaling
- Microtubule and mitosis dynamics
- Protein Degradation and Inhibitors
- Analytical Chemistry and Chromatography
- interferon and immune responses
- Machine Learning in Bioinformatics
- Connexins and lens biology
- Poxvirus research and outbreaks
- Forensic Toxicology and Drug Analysis
German Cancer Research Center
2004-2024
Heidelberg University
2003-2024
German Center for Infection Research
2018-2024
IFOM
2005-2017
University of Oxford
2009-2013
John Radcliffe Hospital
2010
CEA Paris-Saclay
2007
University of Edinburgh
2007
Wellcome Centre for Cell Biology
2007
Commissariat à l'Énergie Atomique et aux Énergies Alternatives
2007
T cell antigen receptor (TCR) and coreceptor ligation is thought to initiate signal transduction by inducing activation of the kinase Lck.Here we showed that catalytically active Lck was present in unstimulated naive cells thymocytes readily detectable these lymphoid organs.In up ∼40% total constitutively activated, part which also phosphorylated on C-terminal inhibitory site.Formation activated independent TCR coreceptors but required catalytic activity its maintenance relied monitoring...
Maintenance of genomic stability depends on the DNA damage response, an extensive signaling network that is activated by lesions such as double-strand breaks (DSBs). The primary activator mammalian DSB response nuclear protein kinase ataxia–telangiectasia, mutated (ATM), which phosphorylates key players in various arms this network. activation and stabilization p53 play a major role are mediated ATM-dependent posttranslational modifications Mdm2, ubiquitin ligase p53. p53's to also...
Human serum is thought to contain key information for diagnostics of human disease. However, no single technology currently nor might ever be available cope with the complexity and dynamic range proteome. We here report a large-scale proteomic study blood using peptide library beads mass spectrometry. Serum proteins are adsorbed onto polymeric coated combinatorial composed millions hexameric baits. Analysis eluates from this (as obtained 3 eluants different strength, able release 99%...
Connexin43 is degraded by the proteasomal as well lysosomal pathway with ubiquitin playing a role in both degradation pathways. So far, no protein ligase has been identified for any of connexins. By using pull-down assays, here we show binding ligase, Nedd4, to C-terminus connexin43. This observation was confirmed vivo coimmunoprecipitation and immunofluorescence, showing colocalization Nedd4 Binding its interaction partners generally carried out WW domains. Our results indicate that...
Abstract Summary: The central proteomics facilities pipeline (CPFP) provides identification, validation, and quantitation of peptides proteins from LC-MS/MS datasets through an easy to use web interface. It is the first analysis targeted specifically at needs core facilities, reducing data load on staff, allowing facility clients easily access work with their data. Identification performed using multiple search engines, output combined validated state-of-the-art techniques for improved...
A missense C1858T single nucleotide polymorphism in the PTPN22 gene recently emerged as a major risk factor for human autoimmunity. encodes lymphoid tyrosine phosphatase (LYP), which forms complex with kinase Csk and is critical negative regulator of signaling through T cell receptor. The results LYP-R620W variation within LYP-Csk interaction motif. LYP-W620 exhibits greatly reduced gain-of-function inhibitor signaling. Here we show that LYP constitutively interacts its substrate Lck...
The SH2 domain–containing leukocyte protein of 76 kD (SLP-76) is a pivotal element the signaling machinery controlling T cell receptor (TCR)-mediated activation. Here, we identify 14-3-3ε and ζ proteins as SLP-76 binding partners. This interaction was induced by TCR ligation required phosphorylation at serine 376. Ribonucleic acid interference in vitro experiments showed that 376 target hematopoietic progenitor kinase 1 (HPK-1). Interestingly, either S376A mutation or HPK-1 knockdown...
In Dhaka, Bangladesh, fresh isolates of Escherichia coli from 197 patients with diarrhea were investigated for production enterotoxin and possession colonization factor antigen (CFA) I or II. Enterotoxigenic E. (ETEC) was isolated 34% the patients, 67 enterotoxin-positive strains, 75% carried CFAs. Among 68 healthy control persons no strains positive both CFA found. The CFAs in general restricted to certain serotypes coli. a subgroup part an ongoing surveillance study, mixed infection seen...
Stimulation of the T cell antigen receptor (TCR) induces formation a phosphorylation-dependent signaling network via multiprotein complexes, whose compositions and dynamics are incompletely understood. Using stable isotope labeling by amino acids in culture (SILAC)-based quantitative proteomics, we investigated kinetics signal propagation after TCR-induced protein tyrosine phosphorylation. We confidently assigned 77 proteins (of 758 identified) as direct or indirect consequence...
Abstract For rational design of therapeutic vaccines, detailed knowledge about target epitopes that are endogenously processed and truly presented on infected or transformed cells is essential. Many potential (viral mutation‐derived), at low abundance. Therefore, direct detection these peptides remains a challenge. This study presents method for the isolation LC‐MS 3 ‐based targeted low‐abundant human leukocyte antigen (HLA) class‐I‐presented from cells. Human papillomavirus (HPV) was used...
Pancreatic ductal adenocarcinoma (PDAC) is generally refractory to immune checkpoint blockade, although patients with genetically unstable tumors can show modest therapeutic benefit. We previously demonstrated the presence of tumor-reactive CD8+ T cells in PDAC samples. Here, we charted tumor-infiltrating cell repertoire by combining single-cell transcriptomics functional testing receptors (TCRs) for reactivity against autologous tumor cells. On basis a comprehensive dataset including 93 and...
The standard strategy for analysis by tandem mass spectrometry of protein phosphorylation at serine or threonine utilizes the neutral loss H3PO4 (= 97.977/z) from proteolytic peptide molecular ions as marker fragmentation. Manual control automatically performed loss-based phosphopeptide identifications is strongly recommended, since these data may contain false-positive results. These are connected to experimental m/z error, competing fragmentation pathways, limitations in interpretation...
Abstract The activation kinetics of MAPK Erk are critical for T cell development and activation. In particular, sustained signaling is required effector functions, such as IL-2 production. Although Raf-1 triggers transient activation, B-Raf implicated in after TCR stimulation. this study, we show that dephosphorylated on its inhibitory serine 365 upon triggering. However, it unknown how coupled to the TCR. Using mass spectrometry, identified protein kinase D–interacting substrate 220 kDa...
Fecal Escherichia coli isolates from 196 patients with watery diarrhea and 68 healthy individuals (controls) were analyzed in Bangladesh immediately after isolation for the presence of colonization factor antigen (CFA) I or II (CFA/I CFA/II, respectively) by a mannose-resistant hemagglutination (MRHA) test six species erythrocytes slide agglutination absorbed CFA/I CFA/II antisera. The CFAs was confirmed immunodiffusion analyses done Sweden. By these methods, it found that 49 69...
Signaling through the T cell receptor (TCR) initiates adaptive immunity and its perturbation may results in autoimmunity. The plasma membrane scaffolding protein LAT acts as a central organizer of TCR signaling machinery to activate many functional pathways. LAT-deficient mice develop an autoimmune syndrome but mechanism this pathology is unknown. In work we have compared global dynamics by MS-based quantitative phosphoproteomics LAT-sufficient LAT-defective Jurkat cells. Surprisingly, found...
Son of Sevenless 1 (SOS1) is a dual guanine nucleotide exchange factor (GEF) that activates the small GTPases RAC and RAS. Although molecular mechanisms RAS GEF catalysis have been unveiled, how SOS1 acquires activity what physio-pathological relevance this much less understood. Here we show tyrosine phosphorylated on Y1196 by ABL. Phosphorylation controls inter-molecular interaction, required to promote nucleotides in vitro for platelet-derived growth (PDGF) activation RAC- RAC-dependent...
Tyrosine-O-sulfated peptides were studied by nanoESI Q-TOF mass spectrometry and found to exhibit an abundant loss of SO3 in positive ion mode under the usually nonfragmenting conditions survey spectrum acquisition. A new strategy for detection tyrosine-O-sulfated total protein digests was designed based on exhaustive product scanning at collision offset typical recording spectra (minimum offset). From these data, neutral scans 80/z precursor ions extracted. The specificity this approach...
A novel highly sensitive strategy is introduced for analysis of tyrosine phosphorylation in previously identified proteins channelling this aim all analytical and sequence information available. Nanoelectrospray high-resolution MS/MS targeted to precalculated m/z values corresponding phosphotyrosine-containing tryptic peptides. Identification these peptides supported by the occurrence phosphotyrosine immonium ion at 216, neutral loss 79.97/z (= HPO3), similarity fragmentation patterns with...
Early downstream responses of T lymphocytes following cell antigen receptor (TCR) activation are mediated by protein complexes that assemble in domains the plasma membrane. Using stable isotope labeling with amino acids culture and mass spectrometry, we quantitatively related proteome αCD3 immunoisolated native TCR signaling membrane to control fragments not engaged signaling. Proteins were sorted according their relative enrichment isolated domains, identifying a complex network is anchored...
The widespread occurrence of the neutral loss one to six amino acid residues as fragments from doubly protonated tryptic peptides is documented for 23 with individual sequences. Neutral acids N-terminus charged results in y-ions, forming a ladder-like series ions [M + 2H](2+) = y(max) (2+), y(max - 1) 2) etc. An internal residue such histidine, proline, lysine or arginine appears favor this type fragmentation, although it was sometimes also observed without structure. For non-tryptic these...
• The AGC protein kinase OXI1 is a key in plant responses to oxidative signals, and important for two burst-mediated processes: basal resistance microbial pathogens root hair growth. To identify possible components of the signalling pathway, phosphoproteomic techniques were used detect alterations abundance phosphorylated proteins peptides an oxi1 null mutant Arabidopsis thaliana. relative was assessed either using two-dimensional gel electrophoresis staining with phosphoprotein stain Pro-Q...
Peptides containing a monoiodo- or diiodo-tyrosine residue (monoiodo-Y, diiodo-Y) were found to generate abundant immonium ions following collision-induced dissociation at m/z 261.97 and 387.87 Da, respectively. These residue-specific marker are between about 140 mDa (monoiodo-Y) 300 (diiodo-Y) mass deficient relative any other peptide fragment of unmodified peptides, qualifying them as highly specific for tyrosine iodination when analyzed by high resolution tandem spectrometry (MS/MS). Two...