1. Presence of Ceramidase Activity in Electronegative LDL
- Author
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Puig, Núria, Rives, Jose, Estruch, Montserrat, Aguilera-Simon, Ana, Rotllan, Noemi, Camacho, Mercedes, Colomé, Núria, Canals, Francesc, Sanchez-Quesada, Jose Luis, Benitez, Sonia, Universitat Autònoma de Barcelona, Institut Català de la Salut, [Puig N, Rives J] Cardiovascular Biochemistry Group, Research Institute of the Hospital de la Santa Creu i Sant Pau (IIB Sant Pau), Barcelona, Spain. Departament de Biologia Molecular i Bioquímica, Universitat Autònoma de Barcelona, Bellaterra, Spain. [Estruch M] Biotech Research and Innovation Centre, Faculty of Health and Medical Sciences, University of Copenhagen BRIC, Copenhagen, Denmark. [Aguilera-Simon A] Vascular Brain Diseases, IIB Sant Pau, Barcelona, Spain. [Rotllan N] Molecular Basis of Cardiovascular Risk, IIB Sant Pau, Barcelona, Spain. CIBER of Diabetes and Related Metabolic Diseases (CIBERDEM), Instituto de Salud Carlos III, Madrid, Spain. [Camacho M] Genomics of Complex Diseases Unit, IIB Sant Pau, Barcelona, Spain. CIBER of Cardiovascular Disease (CIBERCV), Instituto de Salud Carlos III, Madrid, Spain. [Colomé N, Canals F] Proteomics Laboratory, Vall d’Hebron Institute of Oncology (VHIO), Barcelona, Spain, and Vall d'Hebron Barcelona Hospital Campus
- Subjects
compuestos orgánicos::compuestos orgánicos::compuestos orgánicos::aminas::alcoholes amino::esfingosina [COMPUESTOS QUÍMICOS Y DROGAS] ,disciplinas de las ciencias naturales::disciplinas de las ciencias biológicas::bioquímica::proteómica [DISCIPLINAS Y OCUPACIONES] ,Organic Chemistry ,electronegative LDL ,ceramide ,sphingosine ,ceramidase ,sphingomyelinase ,Ceramidase ,General Medicine ,Proteòmica ,Electronegative LDL ,Catalysis ,Computer Science Applications ,Ceramide ,Inorganic Chemistry ,Natural Science Disciplines::Biological Science Disciplines::Biochemistry::Proteomics [DISCIPLINES AND OCCUPATIONS] ,enzimas y coenzimas::enzimas::hidrolasas::amidohidrolasas::ceramidasas [COMPUESTOS QUÍMICOS Y DROGAS] ,Sphingosine ,Sphingomyelinase ,Amines ,Physical and Theoretical Chemistry ,Enzymes and Coenzymes::Enzymes::Hydrolases::Amidohydrolases::Ceramidases [CHEMICALS AND DRUGS] ,Molecular Biology ,Organic Chemicals::Organic Chemicals::Organic Chemicals::Amines::Amino Alcohols::Sphingosine [CHEMICALS AND DRUGS] ,Spectroscopy - Abstract
Ceramide; Sphingomyelinase Ceramida; Esfingomielinasa Ceramida; Esfingomielinasa 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 °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(−). This research was funded by grants PI13/00364, PI16/00471, FIS PI019/00421, and PI20/00334 from the Instituto de Salud Carlos III, Spanish Ministry of Health (co-financed by the European Regional Development Fund). N.P. is the recipient of FI20/00252 from Instituto de Salud Carlos III. This research was supported by CIBER (Consorcio Centro de Investigación Biomédica en Red) (CB07/08/0016), Instituto de Salud Carlos III, and Ministerio de Ciencia e Innovación and Unión Europea—European Regional Development Fund. CIBERDEM (CB07/08/0016) and CIBERCV (CB16/11/00257) are Instituto de Salud Carlos III Projects. A.A.-S. is member of RETICS INVICTUS PLUS (RD16/0019/0010, the Instituto de Salud Carlos III project). N.P., S.B., N.R., and J.L.S.-Q. are members of the Quality Research Group 2017-SGR-1149 from Generalitat de Catalunya and the Group of Vascular Biology of the Spanish Society of Atherosclerosis.
- Published
- 2022
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