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Heterologous Overexpression of Sup35 in E. coli Leads to Both Monomer and Complex States
  • Mingyang Wang,
  • Xiao Wang,
  • Zhenyun Cheng
Mingyang Wang
Clinical Laboratory The First Affiliated Hospital of Zhengzhou University Key Clinical Laboratory of Henan Province Zhengzhou Henan 450052 China

Corresponding Author:[email protected]

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Xiao Wang
Clinical Laboratory The First Affiliated Hospital of Zhengzhou University Key Clinical Laboratory of Henan Province Zhengzhou Henan 450052 China
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Zhenyun Cheng
Clinical Laboratory The First Affiliated Hospital of Zhengzhou University Key Clinical Laboratory of Henan Province Zhengzhou Henan 450052 China
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Abstract

The heterologous overexpression states of prion proteins play a critical role in understanding the mechanisms of prion-related diseases. We report herein the identification of soluble monomer and complex states for a bakers’ yeast prion, Sup35, when expressed in E. coli. Two peaks are apparent with the elution of His-tagged Sup35 by imidazole from a Ni 2+ affinity column. Peak I contains Sup35 in both monomer and aggregated states. Sup35 aggregate is abbreviated as C-aggregate and includes a non-fibril complex comprising Sup35 aggregate-HSP90-Dna K, ATP synthase β unit (chain D), 30S ribosome subunit, and Omp F. The purified monomer and C-aggregate can remain stable for an extended period of time. Peak II contains Sup35 also in both monomer and aggregated (abbreviated as S-aggregate) states, but the aggregated states are caused by the formation of inter-Sup35 disulfide bonds. This study demonstrates that further assembly of Sup35 non-fibril C-aggregate can be interrupted by the chaperone repertoire system in E. coli.
24 Dec 2021Submitted to PROTEINS: Structure, Function, and Bioinformatics
24 Dec 2021Submission Checks Completed
24 Dec 2021Assigned to Editor
31 Dec 2021Reviewer(s) Assigned
07 Jan 2022Review(s) Completed, Editorial Evaluation Pending
11 Jan 2022Editorial Decision: Revise Major
07 Feb 20221st Revision Received
07 Feb 2022Submission Checks Completed
07 Feb 2022Assigned to Editor
07 Feb 2022Reviewer(s) Assigned
11 Feb 2022Review(s) Completed, Editorial Evaluation Pending
11 Feb 2022Editorial Decision: Revise Minor
16 Feb 20222nd Revision Received
16 Feb 2022Submission Checks Completed
16 Feb 2022Assigned to Editor
16 Feb 2022Review(s) Completed, Editorial Evaluation Pending
16 Feb 2022Editorial Decision: Accept
22 Feb 2022Published in Proteins: Structure, Function, and Bioinformatics. 10.1002/prot.26327