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In vitro synthesis of disialoganglioside (G d1α ) from asialo‐G m1 using sialyltransferases in rat liver Golgi vesicles

1994; Wiley; Volume: 221; Issue: 1 Linguagem: Inglês

10.1111/j.1432-1033.1994.tb18772.x

1432-1033

Kazuya I.P.J. Hidari, Ikuo Kawashima, Tadashi Tai, Fuyuhiko Inagaki, Yoshitaka Nagai, Yutaka Sanai,

Caveolin-1 and cellular processes

Two gangliosides were efficiently synthesized from asialo‐G m1 (Galβ1‐3GalNAcβ1‐4Galβ1‐4Glcβ1‐1 Cer) and cytidine 5′‐phosphate‐ N ‐acetylneuraminic acid (CMP‐NeuAc) by using sialyltransferases in rat liver Golgi vesicles in vitro . These gangliosides were rapidly purified by a combination of anion exchange and reverse‐phase column chromatographies. The ganglioside structures were determined by TLC analysis, treatment with a sialidase from Salmonella typhimurium LT2, which specifically hydrolyzes α2‐3 N ‐acetylneuraminic acid (NeuAcα2‐3) linkages, TLC immunostaining, and 1 H‐NMR spectroscopy. One of the gangliosides was identified as G d1α [NeuAcα2‐3Galβ1‐3(NeuAcα2‐6)GalNAcβ1‐4Galβ1‐4Glcβ1‐1 Cer]. The other ganglioside was determined to be G m1b (NeuAcα2‐3Galβ1‐3GalNAcβ1‐4Galβ1‐4Glcβ1‐1 Cer), which has been reported in a previous study [Pohlentz, G., Klein, D., Schmitz, D., Schwarzmann, G., Peter‐Katalinic, J. & Sandhoff, K. (1988) Biol. Chem. Hoppe‐Seyler 369 , 55–63]. Finally, G m1b and G d1α were obtained from asialo‐G m1 as a starting material in 8.1% and 1.2% overall yields, respectively. This study also suggests that the novel synthetic pathway asialo‐G m1 → G m1b → G d1α may exist in rat liver.