Effects of a sphingolipid synthesis inhibitor on membrane transport through the secretory pathway

Anne G. Rosenwald, Carolyn E. Machamer, Carolyn E Machamer

Research output: Contribution to journalArticle

Abstract

We investigated the effects of an inhibitor of sphingolipid biosynthesis, 1-phenyl-2-(decanoyl-amino)-3-morpholino-l-propanol (PDMP), on cells in culture. Two Golgi-associated enzymes were affected by incubation of cells with PDMP. The synthesis of glucosylceramide was inhibited at low concentrations of PDMP (2.5-10 μM), and in the presence of higher concentrations (≥25 μM), synthesis of sphingomyelin was also reduced. Transport of vesicular stomatitis virus G protein through the Golgi complex was progressively retarded by increasing concentrations of PDMP. In the presence of 75 μM PDMP, the half-times of VSV-G protein arrival at the cis, medial, and trans Golgi and the cell surface were increased 1.5-, 2.1-, 2.4-, and 2.8-fold, respectively, compared to control values. Transport of fluorescent sphingolipids, synthesized de novo at the Golgi complex from fluorescent ceramide precursors, to the cell surface was retarded by ∼20% in the presence of 50 μM PDMP and by ∼50% in the presence of 100 μM PDMP. Control experiments demonstrated that PDMP had minimal effects on cell morphology and physiology (including microtubule and endoplasmic reticulum structure, mitochondrial function, and endocytosis). Although incubation of cells with relatively high concentrations of PDMP was required to see the effects on protein and sphingolipid transport, use of a fluorescent analogue of PDMP demonstrated that most cell-associated PDMP was sequestered in lysosomes, while the concentration at the Golgi complex, the site of the target synthetic enzymes, was relatively low. Taken together, these results suggest that transport of proteins and sphingolipids through the secretory pathway may be coupled to sphingolipid synthesis.

Original languageEnglish (US)
Pages (from-to)3581-3590
Number of pages10
JournalBiochemistry®
Volume31
Issue number14
StatePublished - 1992

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Sphingolipids
Secretory Pathway
Membranes
Golgi Apparatus
RV 538
Cells
Glucosylceramides
Cell Physiological Phenomena
1-Propanol
Morpholinos
Sphingomyelins
Ceramides
Biosynthesis
Physiology
Protein Transport
Enzymes
Endocytosis
Lysosomes
GTP-Binding Proteins
Microtubules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Effects of a sphingolipid synthesis inhibitor on membrane transport through the secretory pathway. / Rosenwald, Anne G.; Machamer, Carolyn E.; Machamer, Carolyn E.

In: Biochemistry®, Vol. 31, No. 14, 1992, p. 3581-3590.

Research output: Contribution to journalArticle

Rosenwald, Anne G. ; Machamer, Carolyn E. ; Machamer, Carolyn E. / Effects of a sphingolipid synthesis inhibitor on membrane transport through the secretory pathway. In: Biochemistry®. 1992 ; Vol. 31, No. 14. pp. 3581-3590.
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