Structural Characterization of the Intra- and Inter-Repeat Copper Binding Modes within the N-Terminal Region of “Prion Related Protein” (PrP-rel-2) of Zebrafish

The unique biology of prion proteins (PrPs) allied with the public-health risks posed by prion zoonoses, such as various animal neurodegenerations, has focused much attention on the molecular basis of the controls cross-species and on the similarities between PrPs from different species. Given the c...

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Published inThe journal of physical chemistry. B Vol. 112; no. 47; pp. 15140 - 15150
Main Authors Gaggelli, Elena, Jankowska, Elzbieta, Kozlowski, Henryk, Marcinkowska, Alina, Migliorini, Caterina, Stanczak, Pawel, Valensin, Daniela, Valensin, Gianni
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 27.11.2008
Subjects
Amino Acid Sequence
Animals
B: Biophysical Chemistry
Circular Dichroism
Copper - metabolism
Electron Spin Resonance Spectroscopy
Molecular Sequence Data
Nuclear Magnetic Resonance, Biomolecular
Prions - chemistry
Prions - metabolism
Spectrophotometry, Ultraviolet
Zebrafish
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Summary:The unique biology of prion proteins (PrPs) allied with the public-health risks posed by prion zoonoses, such as various animal neurodegenerations, has focused much attention on the molecular basis of the controls cross-species and on the similarities between PrPs from different species. Given the common feature of PrPs as Cu2+ binding proteins, it appears relevant to compare the impact of Cu2+ on the stability constants and structures of “physiological” complexes. After having comprehensively delineated the interaction of Cu2+ with mammalian and avian PrPs, the stabilities and molecular structures of species generated by Cu2+ interacting with the irregular repeated domain derived from Danio rerio zebrafish PrP-rel-2 were investigated. Copper complexes with different zebrafish PrP-rel-2 fragments were analyzed by potentiometric and spectroscopic techniques. The data were interpreted as to provide evidence of all investigated repeat units selectively binding Cu2+ via the His imidazole(s). The structural models obtained from paramagnetic NMR showed an intra- or inter-copper binding according to the number of the His in the sequence. In comparison to the mammalian and avian cases, the enzymatic function referred to SOD-like activity was shown to be rather faint in the fish PrPs cases.
Bibliography:Table 1s. Potentiometric and spectroscopic data for proton and Cu2+ complexes of zPrP63-70. Metal to ligand ratio = 1:1; [Cu2+] = 0.002 M. Table 2s. 1H chemical shift of zPrP63-70 0.8 mM in D2O at pH 7.3 and 298 K. Table 3s. 13C chemical shift of zPrP63-70 0.8 mM in D2O at pH 7.3 and 298 K. Table 4s. Potentiometric and spectroscopic data for proton and Cu2+ complexes of zPrP63-74. Metal to ligand ratio = 1:1; [Cu2+] = 0.001 M. Table 5s. Potentiometric and spectroscopic data for proton and Cu2+ complexes of zPrP63-80. Metal to ligand ratio = 1:1; [Cu2+] = 0.001 M. Table 6s. Potentiometric and spectroscopic data for proton and Cu2+ complexes of zPrP63-87. Metal to ligand ratio = 1:1; [Cu2+] = 0.001 M. Table 7s. IC50 (μM) values of the Cu2+-peptide complexes in pH 7.4 with respect to the native Cu, Zn-SOD enzyme, and Cu(NO3)2·6H2O. Table 8s. R1p (S–1) and koff (S–1) values for zPrP Cu (II) complexes. Table 9s. Cu2+–H distances (nm) of zPrP63-70 derived from R1p values calculated in the presence of 0.5 Cu2+ equivalents (τM = 46−47 ms). zPrP63-70 0.8 mM in D2O at pH 7.3 and 298 K. Table 10s. Cu2+–H distances (nm) of zPrP63-74 derived from R1p values calculated in presence of 0.4 Cu2+ equivalents (τM = 9 ms). zPrP63-74 0.8 mM in D2O at pH 7.3 and 298 K. This material is available free of charge via the Internet at http://pubs.acs.org.
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ISSN:1520-6106
1520-5207
DOI:10.1021/jp804759q