Journal: PLoS Pathogens

Article Title: Cellular N-myristoyltransferases play a crucial picornavirus genus-specific role in viral assembly, virion maturation, and infectivity

doi: 10.1371/journal.ppat.1007203

Figure Lengend Snippet: (A) Work flow for detection of Alk-12 labeled viral proteins with click reagents. HeLa cells (DDD85646- or DMSO-treated) are infected with CVB3 at MOI of 10. The myristic acid analogue Alk-12 is added 4 h p.i. and incubation continued for 3 h; during this time Alk-12 becomes transferred to viral substrate proteins by metabolic incorporation. Cells are lysed and Alk-12-bearing proteins are ligated to the fluorescent reporter 5-TAMRA-azide by click chemistry, separated by SDS-PAGE and visualized by in-gel fluorescence. (B) DDD85646 at 5 μM blocks incorporation of Alk-12 into CVB3 VP0. The gel subjected to in-gel fluorescence recording was subsequently stained with InstantBlue to visualize total proteins for verifying equal loading.

Article Snippet: Click chemistry grade Na-ascorbate, Tris(3-hydroxypropyltriazolylmethyl)amine (THPTA), Cu 2 SO 4 , and the fluorescent capturing reagents Cy5.5-Alkyne and 5-TAMRA-Azide were from Jena Biosciences.

Techniques: Labeling, Infection, Incubation, SDS Page, Fluorescence, Staining

Journal: Cell reports

Article Title: NLRP3 Cys126 palmitoylation by ZDHHC7 promotes inflammasome activation

doi: 10.1016/j.celrep.2024.114070

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: TAMRA-azide , Lumiprobe , Cat# C7130; CAS: 825651–66-9.

Techniques: Recombinant, Control, Cytotoxicity Assay, Enzyme-linked Immunosorbent Assay, Reverse Transcription, SYBR Green Assay, Western Blot, Expressing, Mutagenesis, Transgenic Assay, Plasmid Preparation, Construct, Software, Microscopy

Journal: STAR Protocols

Article Title: Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis

doi: 10.1016/j.xpro.2022.101462

Figure Lengend Snippet: Schematic illustration of fluorescence labeling and imaging of an endogenous SrfAB-NRPS For in vitro labeling of an endogenous SrfAB-NRPS, Asp-AMS-BPyne was added to bacterial proteomes. The samples were then irradiated with UV light (365 nm), reacted with the 5/6-TAMRA-peg 3 -azide dye under Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAc), and visualized by SDS-PAGE coupled with in-gel fluorescence imaging. For in-cell labeling studies, Bacillus subtilis ATCC 21332 was cultured, collected, and incubated with Asp-AMS-BPyne . After washing, the bacterial cells were UV-irradiated, lysed, treated with the 5/6-TAMRA-peg 3 -azide dye, and analyzed by SDS-PAGE. For fluorescence cell imaging, after photoaffinity crosslinking in living bacterial cells using Asp-AMS-BPyne , the cells were fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X-100, reacted with the 5/6-TAMRA-peg 3 -azide dye under live-cell CuAAC conditions, and visualized by fluorescence microscopy. Reprinted with permission from Ishikawa et al. (2022) .

Article Snippet: 5/6-TAMRA-peg 3 -azide , Jena Bioscience , Cat# CLK-AZ109-1.

Techniques: Fluorescence, Labeling, Imaging, In Vitro, Irradiation, SDS Page, Cell Culture, Incubation, Microscopy

Journal: STAR Protocols

Article Title: Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis

doi: 10.1016/j.xpro.2022.101462

Figure Lengend Snippet: In vitro CuAAC click chemistry mix

Article Snippet: 5/6-TAMRA-peg 3 -azide , Jena Bioscience , Cat# CLK-AZ109-1.

Techniques: In Vitro, Concentration Assay