We stained cell lines with lectins that have been used to recognize the following three different epitopes: (i) WGA recognizes all sialic acids, (ii) MAA recognizes α2,3 sialic acid, and (iii) SNA recognizes α2,6 sialic acid. In this report, we demonstrate that AAV1 and AAV6 use both α2,3 and α2,6 N-linked sialic acids for binding […]
Neu5Ac used to be purified from animal sources such as edible bird’s nest (Martin et al., 1977) or egg yolk (Koketsu et al., 1992). However the low sialic acid content of these materials resulted in a low overall production yield and precluded the development of an economically practical industrial process. Glycosidically bound, but not free, […]
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The resulting weight gain during the last 2 min of raising the venous pressure was divided by the concomitant increment of capillary pressure, and values were normalized to predicted lung weight. The amounts of activated Rac1 or Cdc42 (arbitrary units) are based on the optical density values obtained after subtracting negative control values. A “recombinant […]
In competitive binding assays, 10 mm free synthetic sialic acid (Sigma) was added 30 min before the endocytic agent and left during the assay. These were further purified by size-exclusion chromatography with elution in 50 mm KPi, 200 mm NaCl, pH 7.8, dialyzed against assay buffer (20 mm Tris, 150 mm NaCl, 2 mm tris(2-carboxyethyl)phosphine, […]
Treatment of isolated-perfused lungs with neuraminidase from Vibrio cholerae leads to pulmonary edema. A third key observation in our study is the pattern of alveolar edema observed when isolated-perfused lungs were treated with neuraminidase from Vibrio cholerae. A second key observation in our study is that terminally linked sialic acids are important contributors to endothelial […]
For experiments, PC-12 cells were collected via 0.25% trypsin (Gibco) and seeded for culturing in collagen IV (Sigma-Aldrich) coated 4-chamber slides (Nunc Lab-Tek, Merck). PC-12 cells grew as small clusters in suspension and were maintained in a density 5 × 105 cells/ml in culture flasks coated by 0.1% collagen type IV (Sigma-Aldrich). For testing bioactivity […]
Here, we show that treatment with a sialidase significantly improved the capacity of both immature and mature MDDCs to phagocytose Escherichia coli. Namely, neuraminidase-treated DCs show increased gene transcription of proinflammatory and Th1-promoting cytokines and induce greater proliferative responses of T lymphocytes. Dendritic cells (DCs) play an essential role in immunity against bacteria by phagocytosis […]
Neuraminidases and other reagents were obtained from Sigma-Aldrich. To determine whether sialic acids play a role in endothelial barrier function, cells were treated with neuraminidases to hydrolyze sialic acid moieties. Two components of glycocalyx structure that may play a role in determining function in glycoproteins are 1) the terminal sugar residue(s) on the oligosaccharide chain(s) […]
Both mutants were grown on 2,7-anhydro-Neu5Ac (orange), Neu5Ac (blue), glucose (red), or M9 medium alone (black) in 200-µl microtiter plates. All strains were grown on 2,7-anhydro-Neu5Ac (orange), Neu5Ac (blue), glucose (red), or M9 medium alone (black) in 200-µl microtiter plates. Structure of the sialometabolic nan regulon of E. coli K12 strains and the role of […]