Sphingolipid biosynthesis in cultured neurons. Down-regulation of serine palmitoyltransferase by sphingoid bases.
Addition of exogenous sphingosine homologues (D-erythro configuration) with different alkyl chain lengths (12 and 18 carbon atoms) to the medium of primary cultured cerebellar cells resulted in a decrease of serine palmitoyltransferase activity in a time- and concentration-dependent manner. This enzyme catalyzes the first committed step in sphingolipid biosynthesis. Half-maximal reduction of enzyme activity occurred after a 4-h treatment with 25 microM sphingoid bases. Maximal decrease (approx. 80%) was obtained after treating the cells for 4-8 h with 50 microM long-chain bases. When a biosynthetically inert sphingoid, azidosphingosine (10-50 microM), was fed to the cells, decrease of 3-ketosphinganine formation was much slower, reaching its maximum (approx. 80%) after 24 h. In contrast to D-erythro-sphingosine, L-threo-C18-sphingosine did not yield any decrease of serine palmitoyltransferase activity when added to the cells under identical experimental conditions. Decrease of serine palmitoyltransferase activity was fully reversible after removal of the long-chain bases from the culture medium. Activities of other enzymes of lipid metabolism, ceramide synthase, long-chain acyl-CoA synthase and choline phosphotransferase, were not affected by the addition of sphingoid bases, indicating that the down regulation of serine palmitoyltransferase is quite specific.
Mandon
E C
EC
Institut für Organische Chemie und Biochemie, Universität Bonn, Federal Republic of Germany.
van Echten
G
G
Birk
R
R
Schmidt
R R
RR
Sandhoff
K
K
D000217:Acyltransferases / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D002531:Cerebellum / Q000187:drug effects / Q000378:metabolism*; D003066:Coenzyme A Ligases / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008815:Mice, Inbred Strains; D008861:Microsomes / Q000201:enzymology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D010770:Phosphotransferases / Q000378:metabolism; D051102:Serine C-Palmitoyltransferase; D013107:Sphingolipids / Q000096:biosynthesis* / Q000302:isolation & purification; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D051049:Sphingosine N-Acyltransferase; D017855:Transferases (Other Substituted Phosphate Groups)
D013107:Sphingolipids; D000217:Acyltransferases; D051049:Sphingosine N-Acyltransferase; D051102:Serine C-Palmitoyltransferase; C495088:Sptlc1 protein, mouse; D010770:Phosphotransferases; D017855:Transferases (Other Substituted Phosphate Groups); C019863:ceramide cholinephosphotransferase; D003066:Coenzyme A Ligases; D013110:Sphingosine