Key words: folimycin/concanamycinA/intracellular translocation of glycoproteins/vacuolar-type ATPase/vesicular stomatitis virus/the Golgi apparatus ABSTRACT. Folimycin (concanamycin A) specifically inhibited vacuolar-type ATPase as far as examined. Folimycin blocked excretion of the glycoprotein (G protein) of vesicular stomatitis virus into the medium and, instead, G protein was accumulated intracellularly.The intracellularly accumulated G protein electrophoresed a little faster than mature one. The N-glycan of the G protein was endoglycosidase H-sensitive, and terminal galactose and N-acetylglucosamine were not detected essentially on sequential digestion with exoglycosidases, indicating that processings known to occur in the Golgi apparatus do not take place in the presence of folimycin. The oligosaccharide chain of the G protein was determined to have a composition of Man8GlcNAc2 as analyzed by Bio-Gel P-4 column chromatography and high-performance liquid chromatography following digestion with a-and then with /5-mannosidase. Activities of mannosidase I and glycosyltransferases prepared from baby hamster kidney cells were not inhibited as far as examined, indicating that the incompleteness of the N-glycosidic chain in folimycin-treated cells is not caused by inhibition of processing enzymes. Taken together these observations suggest that folimycin blocks the intracellular translocation of G protein before the step of trimming by mannosidase I which is confined to the cis compartment of the Golgi. The intracellular localization of G protein as revealed by fluorescence microscopy was in good accordance with this assumption.
The effects of four-week raffinose intake (15 g/day) on the fecal microflora and fecal properties were studied in healthy human volunteers. The significantly increasing numbers of Bifidobacterium spp. were observed during the raffinose intake, whereas the numbers of lecithinase-negative Clostridium spp. and bacteroidaceae during the intake were significantly lower than those before and after the intake. The percentage of Bifidobacterium spp. was increased from 11.6-15.5% of the total to 58.2-80.1% of the total during the intake. The fecal pH values during the feeding were lower than those before and after the intake.
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