Effects of Cu2+ on conformational change and aggregation of hPrP180-192 with a V180I mutation of the prion protein

Yuko Sakaguchi; Rina Nakamura; Motomi Konishi; Yusuke Hatakawa; Hidenao Toyoda; Toshifumi Akizawa

doi:10.1016/j.bbrc.2019.05.009

Effects of Cu²⁺ on conformational change and aggregation of hPrP180-192 with a V180I mutation of the prion protein

Biochem Biophys Res Commun. 2019 Jun 30;514(3):798-802. doi: 10.1016/j.bbrc.2019.05.009. Epub 2019 May 10.

Authors

Yuko Sakaguchi¹, Rina Nakamura², Motomi Konishi³, Yusuke Hatakawa⁴, Hidenao Toyoda⁵, Toshifumi Akizawa⁶

Affiliations

¹ Laboratory of Bio-analytical Chemistry, Faculty of Pharmaceutical Sciences, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga, 525-8577, Japan; Department of Pharmacy, Japanese Red Cross Kyoto Daiichi Hospital, 15-749 Honmachi, Higashiyama-ku, Kyoto, Kyoto, 605-0981, Japan. Electronic address: yuko-sakaguchi@kyoto1-jrc.org.
² O-Force Co., Ltd, 3454 Irino Kuroshio-cho, Hata-gun, Kochi, 789-1931, Japan; Laboratory of Pharmacology, School of Medicine, Kochi University, Kohasu, Oko-cho, Nankoku, Kochi, 783-0047, Japan. Electronic address: rinapirika1024@gmail.com.
³ Analytical Chemistry, Pharmaceutical Science, Setsunan University, 45-1 Nagaotoge-cho, Hirakata, Osaka, 573-0101, Japan. Electronic address: motomi@pharm.setsunan.ac.jp.
⁴ Analytical Chemistry, Pharmaceutical Science, Setsunan University, 45-1 Nagaotoge-cho, Hirakata, Osaka, 573-0101, Japan. Electronic address: yusuke10043175@gmail.com.
⁵ Laboratory of Bio-analytical Chemistry, Faculty of Pharmaceutical Sciences, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga, 525-8577, Japan. Electronic address: hidenao@ph.ritsumei.ac.jp.
⁶ O-Force Co., Ltd, 3454 Irino Kuroshio-cho, Hata-gun, Kochi, 789-1931, Japan; Laboratory of Pharmacology, School of Medicine, Kochi University, Kohasu, Oko-cho, Nankoku, Kochi, 783-0047, Japan. Electronic address: momizit0510@gmail.com.

PMID: 31079927
DOI: 10.1016/j.bbrc.2019.05.009

Abstract

Prion diseases are neurodegenerative disorders caused by misfolding of the prion protein (PrP) from a normal cellular protein (PrP^C) to a protease-resistant isoform (PrP^Sc). However, the aggregation mechanism is not entirely understood because of the physical properties of PrP, such as its solubility or aggregation in vitro and conformational or mutation diversity. Recently, we reported the physical and physiological properties of a synthetic fragment peptide. In the present study, we assessed the importance of a point mutation at the C-terminal region of PrP in structural conversion and aggregation and evaluated the physical and physiological properties of the point-mutated human-PrP180-192 V180I (hPrP180-192 V180I) using circular dichroism spectra, high-performance liquid chromatography, Affinix QNμ, and thioflavin-T staining, including the effects of Cu²⁺. The secondary structure of hPrP180-192 V180I changed from a random coil to a β-sheet in Cu²⁺ free buffer. In addition, we observed molecular interactions in hPrP180-192 V180I and aggregation with itself, which were inhibited by Cu²⁺. We conclude that the point mutation in the C-terminal region of PrP, including hPrP180-192 V180I, and Cu²⁺ may play an important role in the conversion of PrP^C to PrP^Sc.

Keywords: Aggregation; Circular dichroism spectra; Cu(2+); HPLC analysis; Intermolecular interaction; Point-mutated hPrP180-192; Prion protein; Structural conversion; Thioflavin-T.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Benzothiazoles / metabolism
Copper / pharmacology*
Humans
Mutation / genetics*
Prion Proteins / chemistry*
Prion Proteins / genetics*
Protein Aggregates*
Protein Structure, Secondary
Time Factors

Substances

Benzothiazoles
Prion Proteins
Protein Aggregates
thioflavin T
Copper