copper-triggered aggregation of ubiquitincopper-triggered聚合的泛素.pdfVIP

Copper-Triggered Aggregation of Ubiquitin 1 1 ` 2 1 3 Fabio Arnesano *, Simone Scintilla , Vincenza Calo , Elena Bonfrate , Chiara Ingrosso , Maurizio 1 4 5 1 Losacco , Teresa Pellegrino , Enrico Rizzarelli , Giovanni Natile 1 Dipartimento Farmaco-Chimico, University of Bari ‘‘A. Moro’’, Bari, Italy, 2 Consorzio Interuniversitario di Ricerca in Chimica dei Metalli nei Sistemi Biologici (CIRCMSB), Bari, Italy, 3 Dipartimento di Chimica, University of Bari ‘‘A. Moro’’, Bari, Italy, 4 National Nanotechnology Laboratory of CNR-INFM and IIT Research Unit, University of Salento, Lecce, Italy, 5 Dipartimento di Scienze Chimiche, University of Catania, Catania, Italy Abstract Neurodegenerative disorders share common features comprising aggregation of misfolded proteins, failure of the ubiquitin- proteasome system, and increased levels of metal ions in the brain. Protein aggregates within affected cells often contain ubiquitin, however no report has focused on the aggregation propensity of this protein. Recently it was shown that copper, differently from zinc, nickel, aluminum, or cadmium, compromises ubiquitin stability and binds to the N-terminus with 0.1 micromolar affinity. This paper addresses the role of copper upon ubiquitin aggregation. In water, incubation with Cu(II) leads to formation of spherical particles that can progress from dimers to larger conglomerates. These spherical oligomers are SDS-resistant and are destroyed upon Cu(II) chelation or reduction to Cu(I). In water/trifluoroethanol (80:20, v/v), a mimic of the local decrease in dielectric constant experienced in proximity to a membrane surface, ubiquitin incubation with Cu(II) causes time-depe