Journal: Cell

Article Title: Serotonin reduction in post-acute sequelae of viral infection

doi: 10.1016/j.cell.2023.09.013

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-mouse IFNAR-1 antibody , Bio X Cell , BE0241; RRID:AB_2687723.

Techniques: Virus, Clinical Proteomics, Recombinant, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Bicinchoninic Acid Protein Assay, SYBR Green Assay, TaqMan Assay, Software, Imaging, Control, Electron Microscopy, Low Protein Binding, Membrane

Journal: Nature

Article Title: Metabolic control of BRISC-SHMT2 assembly regulates immune signaling

doi: 10.1038/s41586-019-1232-1

Figure Lengend Snippet: a) Elution profile of the indicated SHMT2ΔN forms from an S75 10/300 size exclusion chromatography column (single experiment). b) Coomassie-stained SDS-PAGE analysis of the indicated SHMT2ΔN protein preparations (data are representative of two independent experiments). c) BRISC DUB activity against a fluorogenic K63-linked diUb substrate in the presence of the indicated SHMT2ΔN mutants. Data are average +/-SEM of three independent experiments carried out in duplicate. d) BRISC DUB activity against K63-linked hexa-Ub chains in the presence of the SHMT2ΔN or SHMT1 forms. e) Ubiquitylation levels of IFNAR1 after IFNα stimulation in 293T cells overexpressing the indicated Abraxas2 and SHMT2ΔN forms (LL->RR = L211R+L215R). IFNAR1 immunoprecipitation (IP) was performed under denaturing conditions and ubiquitin levels were detected using the vu-1 antibody. Mock IP was performed using a generic rabbit IgG antibody. f) Immunoprecipitation (IP) performed using anti-Flag antibody in MEFs transiently transfected with Flag-HA epitope-tagged SHMT2ΔN or mutants. Immunoblot was performed for Abraxas2 and SHMT2 as indicated. UTF = untransfected cells used as control. g) MEFs overexpressing the indicated SHMT2ΔN or mutants were challenged with LPS and interferon receptor-dependent signal transduction response was assessed by immunoblot for STAT1 phosphorylated at Y701 (pSTAT1). Data shown in d - e are representative of three independent experiments. For gel source data, see .

Article Snippet: IFNAR1 were immunoprecipitated with a corresponding antibody (Bethyl Laboratories A304-290A).

Techniques: Size-exclusion Chromatography, Staining, SDS Page, Activity Assay, Immunoprecipitation, Ubiquitin Proteomics, Transfection, Western Blot, Control, Transduction

Journal: Nature

Article Title: Metabolic control of BRISC-SHMT2 assembly regulates immune signaling

doi: 10.1038/s41586-019-1232-1

Figure Lengend Snippet: K63-linked poly-ubiquitylated interferon receptors (IFNAR1/2) are internalized and degraded via the lysosomal degradation pathway. The BRISC-SHMT2 complex is required for IFNAR1/2 deubiquitylation and receptor stabilization at the membrane. BRISC deubiquitylation of IFNAR1/2 receptors proceeds after SHMT2 displacement leading to sustained interferon signaling. SHMT2 dimer-tetramer transition is regulated by pyridoxal-5’-phosphate (PLP) binding. Only an inactive SHMT2 dimer can interact with BRISC, whereas the active SHMT2 tetramer will sterically clash with BRCC36-Abraxas2 superdimer. Modulation of SHMT2 dimer-tetramer transition by PLP analogues or other binders could represent new ways to inhibit BRISC recruitment to IFNAR1/2 and reduction of interferon signaling.

Article Snippet: IFNAR1 were immunoprecipitated with a corresponding antibody (Bethyl Laboratories A304-290A).

Techniques: Membrane, Binding Assay, Analogues