In addition, we determined by inhibitor (N-benzyl GalNAc)- and cell line-specific (Lec-1) studies that AAV1 and AAV6 require N-linked and not O-linked sialic acid. E. coli can transport and catabolize the common sialic acid, Neu5Ac, as a sole source of carbon and nitrogen but also related sialic acids, N-glycolylneuraminic acid (Neu5Gc) and 3-keto-3-deoxy-d-glycero-d-galactonononic acid (KDN), […]
بایگانی نویسنده: petercedillo0
Different AAV serotypes display distinct tissue tropism, believed to be related to the distribution of their receptors on target cells. The AAV serotype helper plasmids pXR1, pXR2, pXR4, and pXR5, as well as the plasmid pXX6-80 containing the adenovirus helper genes for AAV replication, have been described elsewhere (32, 47). AAV6 helper plasmid pXR6 was […]
ΔOD595 for triplicate experiments is shown: ΔnanT (A) and ΔyjhB (B). ΔOD595 for triplicate experiments is shown: BW25113 (B), ΔyjhC (C), and complemented yjhC (D). Deletion of yjhC resulted in loss of growth on 2,7-anhydro-Neu5Ac but not on Neu5Ac (Fig. 5C), which could be complemented in trans with yjhC (Fig. 5D), suggesting that the gene […]
Neu5Ac (blue), 2,7-anhydro-Neu5Ac (orange), or XY intermediate (gray). B and C, superposition of the 1H,13C HSQC reference spectra of Neu5Ac (blue) and 2,7-anhydro-Neu5Ac (orange) and 1H,13C HSQC reaction mixture at 30 min (where the XY peak is observed to peak, gray). The comparison of the 2D 1H,13C HSQC spectra of the substrate (2,7-anhydro-Neu5Ac), the product […]
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, […]
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 […]