N-acylated serinol is a novel ceramide mimic inducing apoptosis in neuroblastoma cells.
A novel structural analog of ceramide was synthesized by N-acylation of serinol (2-amino-1,3-propanediol) and studied for its effects on glycolipid biosynthesis and cell differentiation of neuroblastoma cells. Incubation with N-palmitoylated serinol (C16-serinol) increased the concentration of endogenous ceramide by 50-80% and caused apoptosis in rapidly dividing low density cells but not in confluent cells. Cell death was not suppressed by simultaneous incubation with phorbol ester, known to antagonize ceramide-induced apoptosis by activation of protein kinase C (PKC). Purification of potential target proteins of C16-serinol was achieved by affinity chromatography of a protein preparation from rat brain on immobilized C16-serinol. A gel activity assay revealed that the eluate from C16-serinol-Sepharose contained three serine/threonine-specific protein kinases with molecular masses of 50, 70, and 95 kDa. The 70-kDa protein was immunostained on a Western blot using a PKCzeta-specific antibody. The purified PKCzeta could be activated directly by C16-serinol in an in vitro phosphorylation assay. Induction of apoptosis in neuroblastoma cells was suppressed by inhibition of PKCzeta with Gö 6983. Our overall results indicate that apoptosis in neuroblastoma cells induced by C16-serinol was at least partially mediated by activation of PKCzeta on condition of ongoing cell division. N-Acylated serinols may thus be useful for induction of apoptosis in mitotic cells and may be of therapeutic potential for treatment of cancer in the nervous system.
Bieberich
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Department of Biochemistry, Medical College of Virginia Campus of Virginia Commonwealth University, Richmond, Virginia 23298-0614, USA. ebieberi@hsc.vcu.edu
Kawaguchi
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T
Yu
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RK
D000818:Animals; D017209:Apoptosis; D001921:Brain / Q000201:enzymology; D002518:Ceramides / Q000494:pharmacology*; D005227:Fatty Acids / Q000494:pharmacology*; D051379:Mice; D018716:Molecular Mimicry; D009417:Nerve Tissue / Q000187:drug effects; D009447:Neuroblastoma; D010169:Palmitic Acids / Q000494:pharmacology; D011412:Propanolamines / Q000494:pharmacology; D011409:Propylene Glycols / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D051381:Rats; D015398:Signal Transduction; D013107:Sphingolipids / Q000378:metabolism; D013229:Stearic Acids / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D000099221:Protein Kinase C zeta
D002518:Ceramides; D005227:Fatty Acids; D010169:Palmitic Acids; D011412:Propanolamines; D011409:Propylene Glycols; D011493:Protein Kinase C; D013107:Sphingolipids; D013229:Stearic Acids; D000099221:Protein Kinase C zeta; C403172:N-palmitoylserinol; C403171:N-stearylserinol