Molecular basis for the protective effects of low-density lipoprotein receptor-related protein 1 (LRP1)-derived peptides against LDL aggregation
Por:
Benitez-Arnaro, A, Pallara, C, Nasarre, L, Rivas-Urbina, A, Benitez, S, Vea, A, Bornachea, O, de Gonzalo-Calvo, D, Serra-Mir, G, Villegas, S, Prades, R, Sanchez-Quesada, JL, Chiva, C, Sabido, E, Tarrago, T, Llorente-Cortes, V
Publicada:
1 jul 2019
Resumen:
Aggregated LDL is the first ligand reported to interact with the cluster II CR9 domain of low-density lipoprotein receptor-related protein 1 (LRP1). In particular, the C-terminal half of domain CR9, comprising the region Gly(1127)-Cys(1140) exclusively recognizes aggregated LDL and it is crucial for aggregated LDL binding. Our aim was to study the effect of the sequence Gly(1127)-Cys(1140) (named peptide LP3 and its retro-enantio version, named peptide DP3) on the structural characteristics of sphingomyelinase- (SMase) and phospholipase 2 (PLA(2))-modified LDL particles. Turbidimetry, gel filtration chromatography (GFC) and transmission electronic microscopy (TEM) analysis showed that LP3 and DP3 peptides strongly inhibited SMase- and PLA(2)-induced LDL aggregation. Nondenaturing polyacrylamide gradient gel electrophoresis (GGE), agarose gel electrophoresis and high-performance thin-layer chromatography (HPTLC) indicated that LP3 and DP3 prevented SMase-induced alterations in LDL particle size, electric charge and phospholipid content, respectively, but not those induced by PLA(2). Western blot analysis showed that LP3 and DP3 counteracted changes in ApoB-100 conformation induced by the two enzymes. LDL proteomics (LDL trypsin digestion followed by mass spectroscopy) and computational modeling methods evidenced that peptides preserve ApoB-100 conformation due to their electrostatic interactions with a basic region of ApoB-100. These results demonstrate that LRP1-derived peptides are protective against LDL aggregation, even in conditions of extreme lipolysis, through their capacity to bind to ApoB-100 regions critical for ApoB-100 conformational preservation. These results suggests that these LRP1(CR9) derived peptides could be promising tools to prevent LDL aggregation induced by the main proteolytic enzymes acting in the arterial intima.
Filiaciones:
Benitez-Arnaro, A:
Hosp Santa Creu & Sant Pau, Biomed Res Inst St Pau IIB St Pau, Grp Lipids & Cardiovasc Pathol, Barcelona, Spain
CSIC, Spanish Natl Res Council, Inst Biomed Res Barcelona IIBB, Barcelona, Spain
Pallara, C:
Iproteos SL, Barcelona Sci Pk PCB, Barcelona, Spain
Nasarre, L:
Hosp Santa Creu & Sant Pau, Biomed Res Inst St Pau IIB St Pau, Grp Lipids & Cardiovasc Pathol, Barcelona, Spain
Rivas-Urbina, A:
Hosp St Pau IIB St Pau, Res Inst, Cardiovasc Biochem Grp, Barcelona, Spain
Benitez, S:
Hosp St Pau IIB St Pau, Res Inst, Cardiovasc Biochem Grp, Barcelona, Spain
Vea, A:
Hosp Santa Creu & Sant Pau, Biomed Res Inst St Pau IIB St Pau, Grp Lipids & Cardiovasc Pathol, Barcelona, Spain
Bornachea, O:
Hosp Santa Creu & Sant Pau, Biomed Res Inst St Pau IIB St Pau, Grp Lipids & Cardiovasc Pathol, Barcelona, Spain
CSIC, Spanish Natl Res Council, Inst Biomed Res Barcelona IIBB, Barcelona, Spain
de Gonzalo-Calvo, D:
Hosp Santa Creu & Sant Pau, Biomed Res Inst St Pau IIB St Pau, Grp Lipids & Cardiovasc Pathol, Barcelona, Spain
CSIC, Spanish Natl Res Council, Inst Biomed Res Barcelona IIBB, Barcelona, Spain
CIBER Enfermedades Cardiovasculares CIBERCV, Madrid, Spain
Serra-Mir, G:
Univ Autonoma Barcelona, Fac Biociencies, Dept Bioquim & Biol Mol, Prot Design & Immunotherapy Grp, Barcelona, Spain
Villegas, S:
Univ Autonoma Barcelona, Fac Biociencies, Dept Bioquim & Biol Mol, Prot Design & Immunotherapy Grp, Barcelona, Spain
Prades, R:
Iproteos SL, Barcelona Sci Pk PCB, Barcelona, Spain
Sanchez-Quesada, JL:
Hosp St Pau IIB St Pau, Res Inst, Cardiovasc Biochem Grp, Barcelona, Spain
CIBER Diabet & Enfermedades Metabol Asociadas CIB, Madrid, Spain
Chiva, C:
Barcelona Inst Sci & Technol, Ctr Regulacio Genom, Proteom Unit, Barcelona, Spain
Univ Pompeu Fabra, Barcelona, Spain
Sabido, E:
Barcelona Inst Sci & Technol, Ctr Regulacio Genom, Proteom Unit, Barcelona, Spain
Univ Pompeu Fabra, Barcelona, Spain
Tarrago, T:
Iproteos SL, Barcelona Sci Pk PCB, Barcelona, Spain
Llorente-Cortes, V:
Hosp Santa Creu & Sant Pau, Biomed Res Inst St Pau IIB St Pau, Grp Lipids & Cardiovasc Pathol, Barcelona, Spain
CSIC, Spanish Natl Res Council, Inst Biomed Res Barcelona IIBB, Barcelona, Spain
CIBER Enfermedades Cardiovasculares CIBERCV, Madrid, Spain
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