Redox induces diverse effects on recombinant human wild-type PrP and mutated PrP with inserted or deleted octarepeats

Shi,Qi; Chen,Cao; Zhang,Bao‑Yun; Zhou,Wei; Xiao,Kang; Dong,Xiao‑Ping

doi:10.3892/ijmm.2018.3441

Redox induces diverse effects on recombinant human wild-type PrP and mutated PrP with inserted or deleted octarepeats

Authors:
- Qi Shi
- Cao Chen
- Bao‑Yun Zhang
- Wei Zhou
- Kang Xiao
- Xiao‑Ping Dong
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Affiliations: State Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases (Zhejiang University, Hangzhou), National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, P.R. China
Published online on: January 30, 2018 https://doi.org/10.3892/ijmm.2018.3441
Pages: 2413-2419

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Abstract

Normal prion protein (PrP) contains two cysteines at amino acids 179 and 214, which may form intra‑ and interpeptide disulfide bonds. To determine the possible effects of this disulfide bridge on the biochemical features of PrP, prokaryotic recombinant human wild‑type PrP (PG5), and mutated PrPs with seven extra octarepeats (PG12) or with all five octarepeats removed (PG0), were subjected to redox in vitro. Sedimentation assays revealed a large portion of aggregation in redox‑treated PG5, but not in PG0 and PG12. Circular dichroism analysis detected increased β‑sheet and decreased α‑helix in PG5 subjected to redox, increased random‑coil and decreased β‑sheet in PG0, and increased random‑coil, but limited changes to β‑sheet content, in PG12. Thioflavin T fluorescence tests indicated that fluorescent value was increased in PG5 subjected to redox. In addition, proteinase K (PK) digestions indicated that PK resistance was stronger in PG12 and PG0 compared with in PG5; redox enhanced the PK resistance of all three PrP constructs, particularly PG0 and PG12. These data indicated that formation of a disulfide bond induces marked alterations in the secondary structure and biochemical characteristics of PrP. In addition, the octarepeat region within the PrP peptide markedly influences the effects of redox on the biochemical phenotypes of PrP, thus highlighting the importance of the number of octarepeats in the biological functions of PrP.

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