Project description:Label free quantititative phosphoproteomics analysis following TiO2 enrichment, nanoscale capillary chromatography and high resolution tandem mass spectrometry.
Project description:Human neutrophils and neutrophil-like cells were differentiated from HL-60 cells. Cells were treated with glucocorticoids, dexamethasone or prednisolone, for proteome and phosphoproteome analysis.
Project description:We applied the ultra-fast proteome profiling workflow to tissues collected from NSG mice treated with PBS (vehicle control), 20,000 U/kg ASNaseWT (Spectrila®), or 20,000 U/kg ASNaseQ59L for 0, 1, 3, 5, and 8 d. At each time point, mice were euthanized, and 13 tissues were collected, including subcutaneous white adipose tissue (SQWAT), gonadal adipose tissue (GAT), lung, spleen, stomach, small and large intestine, liver, kidney, heart, leg muscle, brain, and bone marrow. A total of 507 collected tissue samples were processed for quantitative proteome profiling.
Project description:This study aims to investigate the protein expression profiles in a murine model of dextran sulfate sodium (DSS)-induced colitis using advanced Astral-DIA quantitative proteomics technology. A total of 12 colon tissue samples were analyzed, including 6 from healthy control mice and 6 from DSS-treated mice with induced colitis. Experimental Design Species: Mus musculus (C57BL/6 strain). Tissue Source: Colon tissues were dissected, snap-frozen in liquid nitrogen, and homogenized to extract proteins. Groups: Control Group: Healthy mice without intervention. DSS Group: Mice subjected to 2.5% DSS administration for 7 days to induce colitis, validated by histopathological assessment.
Project description:Toll/interleukin-1 receptor (TIR) domain proteins are immune signaling components and function as NAD+-cleaving enzymes to activate defense responses. Activation of TIRs represses growth and drives cell death in plants and promotes axon degeneration in animals, but how plant TIRs are repressed remains unclear. Here, we show that TIR NADase activity requires a conserved serine residue spatially close to the catalytic glutamate. The plant Ca2+-dependent protein kinases (CPKs), the mammalian Ca2+/calmodulin-dependent protein kinase II delta (CAMK2D) and TANK binding kinase 1 (TBK1) phosphorylate TIR domains at this conserved serine, which blocks TIR NADase activities and functions and thus maintains growth in plants and suppresses SARM1 TIR signaling in animals, respectively. Our findings define a fundamental molecular mechanism by which phosphorylation at a conserved serine residue blocks TIR signaling to balance growth and defense trade-offs.
Project description:Lysosomes are implicated in a wide spectrum of human diseases including monogenic lysosomal storage disorders (LSDs), age-associated neurodegeneration and cancer. Profiling lysosomal content using tag-based lysosomal immunopurification (LysoIP) in cell and animal models allowed major discoveries in the field, however, studying lysosomal dysfunction in human patients remains a challenge. Here, we report the development of the tagless LysoIP method to enable rapid enrichment of lysosomes, via immunoisolation, using the endogenous integral lysosomal membrane protein TMEM192, directly from clinical samples and human cells. Isolated lysosomes are intact and suitable for subsequent multimodal omics analyses. To validate the utility of our approach, we employed the tagless LysoIP to enrich lysosomes from peripheral blood mononuclear cells (PBMCs) derived from fresh blood of patients with CLN3 Batten disease, a neurodegenerative LSD. Altogether, the tagless LysoIP provides a framework to study native lysosomes from patient samples, identify novel biomarkers and discover human-relevant disease mechanisms.
Project description:Glucocorticoids (GCs) are corticosteroid hormones that are commonly used for treating systemic inflammatory diseases and acute infections. Immunosuppressive effects of GCs have been studied in many cell types, particularly macrophages and T cells. Despite the importance and abundance of neutrophils in the human immune system, GC responses remain understudied in neutrophils. Here, we performed quantitative mass spectrometry (MS)-based proteomics of primary neutrophils to acquire phosphoproteome.