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Characterization of a Saccharomyces cerevisiae mutant defective in inositol sphingolipid synthesis

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:7246738

A mutant of S. cerevisiae, auxotrophic for the sphingolipid long chain bases sphinganine or 4-hydroxysphinganine has been further characterized. The mutant is defective in the first step in the synthesis of sphingolipid long chain bases, the formation of 3-keto-sphinganine from palmitoyl CoA and L-serine. Crude membranes from wild type cells exhibit a palmitoyl CoA dependent conversion of (/sup 3/H)serine to a labeled product with the properties of 3-ketosphinganine, whereas membranes from the mutant exhibit negligible activity. As expected from this result, they authors find that 3-ketosphinganine supports growth of the mutant as effectively as erythro-sphinganine. To study the sequelae of long chain base deprivation in the mutant required a method to effectively remove lipophilic long chain bases from the cultured cells. They have found that addition of ..cap alpha..-cyclodextrin to the culture medium evidently binds sphinganine tightly enough such that within an hour they observe inhibition of growth (turbidity, cell count) and incorporation of (/sup 3/H)inositol into sphingolipids. Such treatment also results in a loss of cell viability, a long chain base less death possibly similar to the inositol-less death observed in inositol auxotrophs of yeast.

Research Organization:
Univ. of Kentucky, Lexington
OSTI ID:
7246738
Report Number(s):
CONF-8606151-
Journal Information:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:6; ISSN FEPRA
Country of Publication:
United States
Language:
English