This study constitutes the first report revealing the participation of prominin-1 in glucose metabolism and cytoskeleton alteration. Upon stimulation with high glucose, the expression of prominin-1 was up-regulated with concomitant down-regulation of its phosphorylation. Prominin-1 mutated at its phosphorylation site inflicted a global change in the expression of several genes associated with glucose metabolism in L6 myotube cells. Further, the over-expression of prominin-1 promoted glucose uptake in these cells. Prominin-1 undergoes sustained repression of its expression during confluence and differentiation of L6 myotube cells. The expression of prominin-1 in the MDCK cell modulated cell morphology and promoted cellular migration. These data imply that prominin-1 is involved in glucose metabolism and may regulate cellular glucose through its effect on cytoskeleton.