Presence of Ceramidase Activity in Electronegative LDL
Por:
Puig N., Rives J., Estruch M., Aguilera-Simon A., Rotllan N., Camacho M., Colomé N., Canals F., Sánchez-Quesada J.L., Benitez S.
Publicada:
1 ene 2023
Resumen:
Electronegative low-density lipoprotein (LDL(-)) is a minor modified fraction of human plasma LDL with several atherogenic properties. Among them is increased bioactive lipid mediator content, such as lysophosphatidylcholine (LPC), non-esterified fatty acids (NEFA), ceramide (Cer), and sphingosine (Sph), which are related to the presence of some phospholipolytic activities, including platelet-activating factor acetylhydrolase (PAF-AH), phospholipase C (PLC), and sphingomyelinase (SMase), in LDL(-). However, these enzymes' activities do not explain the increased Sph content, which typically derives from Cer degradation. In the present study, we analyzed the putative presence of ceramidase (CDase) activity, which could explain the increased Sph content. Thin layer chromatography (TLC) and lipidomic analysis showed that Cer, Sph, and NEFA spontaneously increased in LDL(-) incubated alone at 37 degrees C, in contrast with native LDL(+). An inhibitor of neutral CDase prevented the formation of Sph and, in turn, increased Cer content in LDL(-). In addition, LDL(-) efficiently degraded fluorescently labeled Cer (NBD-Cer) to form Sph and NEFA. These observations defend the existence of the CDase-like activity's association with LDL(-). However, neither the proteomic analysis nor the Western blot detected the presence of an enzyme with known CDase activity. Further studies are thus warranted to define the origin of the CDase-like activity detected in LDL(-).
Filiaciones:
Puig N.:
Hosp Santa Creu & St Pau IIB St Pau, Cardiovasc Biochem Grp, Res Inst, Barcelona 08041, Spain
Univ Autonoma Barcelona, Biochem & Mol Biol Dept, Cerdanyola Del Valles 08193, Spain
Rives J.:
Hosp Santa Creu & St Pau IIB St Pau, Cardiovasc Biochem Grp, Res Inst, Barcelona 08041, Spain
Univ Autonoma Barcelona, Biochem & Mol Biol Dept, Cerdanyola Del Valles 08193, Spain
Estruch M.:
Univ Copenhagen BRIC, Fac Hlth & Med Sci, Biotech Res & Innovat Ctr, DK-1350 Copenhagen, Denmark
Aguilera-Simon A.:
IIB St Pau, Vasc Brain Dis, Barcelona 08041, Spain
Rotllan N.:
IIB St Pau, Mol Basis Cardiovasc Risk, Barcelona 08041, Spain
Inst Salud Carlos III, CIBER Diabet & Related Metab Dis CIBERDEM, Madrid 28029, Spain
Camacho M.:
IIB St Pau, Genom Complex Dis Unit, Barcelona 08041, Spain
Inst Salud Carlos III, CIBER Cardiovasc Dis CIBERCV, Madrid 28029, Spain
Colomé N.:
Vall Hebron Inst Oncol VHIO, Prote Lab, Barcelona 08035, Spain
Canals F.:
Vall Hebron Inst Oncol VHIO, Prote Lab, Barcelona 08035, Spain
Sánchez-Quesada J.L.:
Hosp Santa Creu & St Pau IIB St Pau, Cardiovasc Biochem Grp, Res Inst, Barcelona 08041, Spain
Inst Salud Carlos III, CIBER Diabet & Related Metab Dis CIBERDEM, Madrid 28029, Spain
Benitez S.:
Hosp Santa Creu & St Pau IIB St Pau, Cardiovasc Biochem Grp, Res Inst, Barcelona 08041, Spain
Inst Salud Carlos III, CIBER Diabet & Related Metab Dis CIBERDEM, Madrid 28029, Spain
Green Accepted, gold, All Open Access, Gold, Green
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