Journal: ACS chemical biology

Article Title: GalNAc-Tyrosine Is a Ligand of Plant Lectins, Antibodies, and Human and Murine Macrophage Galactose-Type Lectins

doi: 10.1021/acschembio.7b00471

Figure Lengend Snippet: Recombinant hMGL binding of the natural beta-amyloid peptide containing GalNAc-Tyr epitope. Recombinant hMGL protein bound the glycosylated Aβ1–15 neoglycoprotein (6/BSA) with a half maximal binding of 25 nM in an ELISA-based assay. Some binding to the nonglycosylated Aβ1–15 neoglycoprotein (6/BSA) was observed, albeit only at a very high concentration. The fluorescence signal values are the means of three independent experiments. Error bars signify the standard deviation.

Article Snippet: In vitro capture of GalNAc-Tyr-BSA neoglycoprotein ligand by bone marrow derived mouse dendritic cells (mBMDCs; Astarte Biologics, Bothell, WA) was examined by flow cytometry.

Techniques: Recombinant, Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Fluorescence, Standard Deviation

Journal: ACS chemical biology

Article Title: GalNAc-Tyrosine Is a Ligand of Plant Lectins, Antibodies, and Human and Murine Macrophage Galactose-Type Lectins

doi: 10.1021/acschembio.7b00471

Figure Lengend Snippet: GalNAc-Tyr neoglycoprotein selectively bound via MGL-2 receptor found on mBMDCs. mBMDCs from female C57BL/6 mice were incubated at 37 °C with 10 μg/mL of CF640R-labeled BSA, peptide-BSA (compound 12-BSA), or GalNAc-Tyr-BSA (compound 10-BSA). (A) MGL-2+ mBMDCs internalized GalNAc-Tyr neoglycoprotein in vitro. Specificity was determined in the presence of Ca2+ chelator EDTA or GalNAc monosacharide. Preincubation with EDTA produced a 72% decrease in positive cells (p < 0.005), and preincubation with 100 mM GalNAc produced an 81% decrease in positive cells (p < 0.005). (B) Mean fluorescence intensity (MFI) is shown for the subset of cells that were positive in each experiment. For those cells that were positive, the MFI was significantly higher for GalNAc-Tyr than the nonglycosylated peptide or the experiments carried out in the presence of EDTA or GalNAc. Data assessing % binding and MFI of positive cells are expressed as the mean ± SD of three independent experiments. Student’s t test was used to assess statistical differences between groups.

Article Snippet: In vitro capture of GalNAc-Tyr-BSA neoglycoprotein ligand by bone marrow derived mouse dendritic cells (mBMDCs; Astarte Biologics, Bothell, WA) was examined by flow cytometry.

Techniques: Incubation, Labeling, In Vitro, Produced, Fluorescence, Binding Assay