title: New Insight in Copper-Ion Binding to Human Islet Amyloid: The Contribution of Metal-Complex Speciation To Reveal the Polypeptide Toxicity.
abstract: Type-2 diabetes (T2D) is considered to be a potential threat on a global level. Recently, T2D has been listed as a misfolding disease, such as Alzheimer's and Parkinson's diseases. Human islet amyloid polypeptide (hIAPP) is a molecule cosecreted in pancreatic β cells and represents the main constituent of an aggregated amyloid found in individuals affected by T2D. The trace-element serum level is significantly influenced during the development of diabetes. In particular, the dys-homeostasis of Cu(2+) ions may adversely affect the course of the disease. Conflicting results have been reported on the protective role played by complex species formed by Cu(2+) ions with hIAPP or its peptide fragments in vitro. The histidine (His) residue at position 18 represents the main binding site for the metal ion, but contrasting results have been reported on other residues involved in metal-ion coordination, in particular those toward the N or C terminus. Sequences that encompass regions 17-29 and 14-22 were used to discriminate between the two models of the hIAPP coordination mode. Due to poor solubility in water, poly(ethylene glycol) (PEG) derivatives were synthesized. A peptide fragment that encompasses the 17-29 region of rat amylin (rIAPP) in which the arginine residue at position 18 was substituted by a histidine residue was also obtained to assess that the PEG moiety does not alter the peptide secondary structure. The complex species formed by Cu(2+) ions with Ac-PEG-hIAPP(17-29)-NH2 , Ac-rIAPP(17-29)R18H-NH2 , and Ac-PEG-hIAPP(14-22)-NH2 were studied by using potentiometric titrations coupled with spectroscopic methods (UV/Vis, circular dichroism, and EPR). The combined thermodynamic and spectroscopic approach allowed us to demonstrate that hIAPP is able to bind Cu(2+) ions starting from the His18 imidazole nitrogen atom toward the N-terminus domain. The stability constants of copper(II) complexes with Ac-PEG-hIAPP(14-22)-NH2 were used to simulate the different experimental conditions under which aggregate formation and oxidative stress of hIAPP has been reported. Speciation unveils: 1) the protective role played by increased amounts of Cu(2+) ions on the hIAPP fibrillary aggregation, 2) the effect of adventitious trace amounts of Cu(2+) ions present in phosphate-buffered saline (PBS), and 3) a reducing fluorogenic probe on H2 O2 production attributed to the polypeptide alone.
authors: Consiglio Nazionale delle Ricerche, Istituto di Biostrutture e Bioimmagini, Via P. Gaifami 18, 95126, Catania, Italy. leotony@unict.it.
authors: Antonio
authors: A
authors: Magrì
authors: Dipartimento di Farmacia, Università di Pisa, Via Bonanno Pisano, 6, 56126, Pisa, Italy. lamendola@farm.unipi.it.
authors: Diego
authors: D
authors: La Mendola
authors: Dipartimento di Scienze Biomediche e Biotecnologiche, Università degli Studi di Catania, Viale A. Doria 6, 95125, Catania, Italy.
authors: Vincenzo Giuseppe
authors: VG
authors: Nicoletti
authors: Consiglio Nazionale delle Ricerche, Istituto di Biostrutture e Bioimmagini, Via P. Gaifami 18, 95126, Catania, Italy.
authors: Giuseppe
authors: G
authors: Pappalardo
authors: Consiglio Nazionale delle Ricerche, Istituto di Biostrutture e Bioimmagini, Via P. Gaifami 18, 95126, Catania, Italy.|Dipartimento di Scienze Chimiche, Università degli Studi di Catania, Viale A. Doria 6, 95125, Catania, Italy.
authors: Enrico
authors: E
authors: Rizzarelli
mesh_terms: D000818:Animals; D001665:Binding Sites; D056831:Coordination Complexes / Q000737:chemistry* / Q000378:metabolism; D003300:Copper / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry; D006801:Humans; D058228:Islet Amyloid Polypeptide / Q000737:chemistry* / Q000633:toxicity; D011092:Polyethylene Glycols / Q000737:chemistry; D011485:Protein Binding; D017433:Protein Structure, Secondary; D051381:Rats; D013816:Thermodynamics
chemical_list: D056831:Coordination Complexes; D058228:Islet Amyloid Polypeptide; D011092:Polyethylene Glycols; D006639:Histidine; D003300:Copper
keywords: copper; cytotoxicity; diabetes; peptides; potentiometry