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J. Biol. Chem., Vol. 275, Issue 22, 16484-16489, June 2, 2000
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Sphingomyelinase Activity Associated with Human Plasma Low Density Lipoprotein
POSSIBLE FUNCTIONAL IMPLICATIONS*

Juha M. HolopainenDagger , Oula Penate Medina, Antti J. Metso, and Paavo K. J. Kinnunen§

From the Helsinki Biophysics and Biomembrane Group, Department of Medical Chemistry, Institute of Biomedicine, University of Helsinki, Helsinki FIN-00014, Finland

Isolated human plasma low density lipoprotein (LDL) was observed to possess sphingomyelinase activity. Accordingly, the formation of ceramide was catalyzed by LDL at 37 °C using tertiary liposomes composed of sphingomyelin (mole fraction (x) = 0.2), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (x = 0.7), 1,2-dimyristoyl-sn-glycero-3-phospho-rac-glycerol (x = 0.1), and either the fluorescent sphingomyelin analog Bodipy-sphingomyelin or [14C]sphingomyelin as substrates. However, this activity was not present in either very low density lipoprotein or the high density lipoprotein subfractions HDL2 and HDL3. Oxidation of LDL abrogated its sphingomyelinase activity. Aggregation of the liposomes upon incubation with LDL was evident from the light scattering measurements. Microinjection of LDL to the surface of giant liposomes composed of 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), N-palmitoyl-D-sphingomyelin (C16:0-sphingomyelin), and Bodipy-sphingomyelin as a fluorescent tracer (0.75:- 0.20:0.05, respectively) revealed the induction of vectorial budding of vesicles, resembling endocytosis.

* This work was supported in part by the Finnish State Medical Research Council and Biocentrum Helsinki.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Supported by the M.D./Ph.D. program of the University of Helsinki and the Finnish Medical Foundation.

§ To whom correspondence should be addressed: Institute of Biomedicine, Dept. of Medical Chemistry, P. O. Box 8 (Siltavuorenpenger 10A), University of Helsinki, Helsinki FIN-00014, Finland. Tel.: 358-9-1918237; Fax: 358-9-1918276; E-mail: Paavo.Kinnunen@Helsinki.fi.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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