학술논문
Ubiquitin binds the amyloid β peptide and interferes with its clearance pathways.
Document Type
Academic Journal
Author
Bellia F; Consiglio Nazionale delle Ricerche , Istituto di Biostrutture e Bioimmagini , Via P. Gaifami 18 , 95126 Catania , Italy . Email: danilo.milardi@cnr.it.; Lanza V; Consiglio Nazionale delle Ricerche , Istituto di Biostrutture e Bioimmagini , Via P. Gaifami 18 , 95126 Catania , Italy . Email: danilo.milardi@cnr.it.; García-Viñuales S; Consiglio Nazionale delle Ricerche , Istituto di Biostrutture e Bioimmagini , Via P. Gaifami 18 , 95126 Catania , Italy . Email: danilo.milardi@cnr.it.; Ahmed IMM; Consiglio Nazionale delle Ricerche , Istituto di Biostrutture e Bioimmagini , Via P. Gaifami 18 , 95126 Catania , Italy . Email: danilo.milardi@cnr.it.; Pietropaolo A; Dipartimento di Scienze della Salute , Università degli Studi Magna Graecia di Catanzaro , Viale Europa , 88100 , Catanzaro , Italy.; Iacobucci C; Department of Pharmaceutical Chemistry & Bioanalytics , Institute of Pharmacy , Martin Luther University Halle-Wittenberg , 06120 Halle/Saale , Germany.; Malgieri G; Department of Environmental , Biological and Pharmaceutical Sciences and Technologies , University of Campania 'Luigi Vanvitelli' , Via Vivaldi 43 , 81100 Caserta , Italy.; D'Abrosca G; Department of Environmental , Biological and Pharmaceutical Sciences and Technologies , University of Campania 'Luigi Vanvitelli' , Via Vivaldi 43 , 81100 Caserta , Italy.; Fattorusso R; Department of Environmental , Biological and Pharmaceutical Sciences and Technologies , University of Campania 'Luigi Vanvitelli' , Via Vivaldi 43 , 81100 Caserta , Italy.; Nicoletti VG; Dipartimento di Scienze Biomediche e Biotecnologiche (BIOMETEC) , sez. Biochimica medica , Università di Catania , Via Santa Sofia 97 , 95124 Catania , Italy.; Sbardella D; Dipartimento di Scienze Cliniche e Medicina Traslazionale , Università di Roma Tor Vergata , Via Montpellier 1 , 00133 , Roma , Italy.; Tundo GR; Dipartimento di Scienze Cliniche e Medicina Traslazionale , Università di Roma Tor Vergata , Via Montpellier 1 , 00133 , Roma , Italy.; Coletta M; Dipartimento di Scienze Cliniche e Medicina Traslazionale , Università di Roma Tor Vergata , Via Montpellier 1 , 00133 , Roma , Italy.; Pirone L; Consiglio Nazionale delle Ricerche , Istituto di Biostrutture e Bioimmagini , Via Mezzocannone, 16 , Naples I-80134 , Italy.; Pedone E; Consiglio Nazionale delle Ricerche , Istituto di Biostrutture e Bioimmagini , Via Mezzocannone, 16 , Naples I-80134 , Italy.; Calcagno D; Dipartimento di Scienze Chimiche , Università di Catania , V.le Andrea Doria 6 , 95125 Catania , Italy . Email: grassog@unict.it.; Grasso G; Dipartimento di Scienze Chimiche , Università di Catania , V.le Andrea Doria 6 , 95125 Catania , Italy . Email: grassog@unict.it.; Milardi D; Consiglio Nazionale delle Ricerche , Istituto di Biostrutture e Bioimmagini , Via P. Gaifami 18 , 95126 Catania , Italy . Email: danilo.milardi@cnr.it.
Source
Publisher: Royal Society of Chemistry Country of Publication: England NLM ID: 101545951 Publication Model: eCollection Cited Medium: Print ISSN: 2041-6520 (Print) Linking ISSN: 20416520 NLM ISO Abbreviation: Chem Sci Subsets: PubMed not MEDLINE
Subject
Language
English
ISSN
2041-6520
Abstract
Several lines of evidence point to a compromised proteostasis associated with a reduction of the Ubiquitin Proteasome System (UPS) activity in patients affected by Alzheimer's Disease (AD) and suggest that the amyloid β peptide (Aβ) is an important player in the game. Inspired also by many reports, underlining the presence of ubiquitin (Ub) in the amyloid plaques of AD brains, here we set out to test whether Ub may bind the Aβ peptide and have any effect on its clearance pathways. By using an integrated array of MALDI-TOF/UPLC-HRMS, fluorescence, NMR, SPR, Microscale Thermophoresis (MST) and molecular dynamics studies, we consistently demonstrated that Aβ40 binds Ub with a 1 : 1 stoichiometry and K d in the high micromolar range. In particular, we show that the N-terminal domain of the Aβ peptide (through residues D1, E3 and R5) interacts with the C-terminal tail of Ub (involving residues K63 and E64), inducing the central region of Aβ ( 14 HQKLVFFAEDVGSNK 28 ) to adopt a mixed α-helix/β-turn structure. ELISA assays, carried out in neuroblastoma cell lysates, suggest that Aβ competitively binds Ub also in the presence of the entire pool of cytosolic Ub binding proteins. Ub-bound Aβ has a lower tendency to aggregate into amyloid-like fibrils and is more slowly degraded by the Insulin Degrading Enzyme (IDE). Finally, we observe that the water soluble fragment Aβ1-16 significantly inhibits Ub chain growth reactions. These results evidence how the non-covalent interaction between Aβ peptides and Ub may have relevant effects on the regulation of the upstream events of the UPS and pave the way to future in vivo studies addressing the role played by Aβ peptide in the malfunction of proteome maintenance occurring in AD.