Expression, purification, and characterization of a membrane-associated cyclic oligo-adenylate degrader from Sulfolobus islandicus .

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  • Author(s): Zhao R;Zhao R; Yang Y; Yang Y; Yang K; Yang K; Han W; Han W
  • Source:
    STAR protocols [STAR Protoc] 2021 Jan 25; Vol. 2 (1), pp. 100299. Date of Electronic Publication: 2021 Jan 25 (Print Publication: 2021).
  • Publication Type:
    Journal Article; Research Support, Non-U.S. Gov't
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Cell Press Country of Publication: United States NLM ID: 101769501 Publication Model: eCollection Cited Medium: Internet ISSN: 2666-1667 (Electronic) Linking ISSN: 26661667 NLM ISO Abbreviation: STAR Protoc
    • Publication Information:
      Original Publication: [Cambridge, MA] : Cell Press, [2020]-
    • Subject Terms:
      Archaeal Proteins*/biosynthesis ; Archaeal Proteins*/chemistry ; Archaeal Proteins*/genetics ; Archaeal Proteins*/isolation & purification ; CRISPR-Cas Systems* ; Gene Expression*; Cell Membrane/*enzymology ; Sulfolobus/*enzymology; Cell Membrane/genetics ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Recombinant Proteins/isolation & purification ; Sulfolobus/genetics
    • Abstract:
      Type III CRISPR-cas systems initiate cyclic oligo-adenylate (cOA) signaling to initiate immune response. Previously, we identified that a membrane-associated DHH-DHHA1 family protein from Sulfolobus islandicus efficiently degrades cOA. Here, we provide detailed protocols for expression and purification of the protein from its native host and a cOA degradation assay with the purified enzyme. The methodology should be of interest for researchers studying Sulfolobus , membrane-associated proteins, or type III CRISPR-cas systems. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2020).
      Competing Interests: The authors declare no competing interests.
      (© 2021 The Author(s).)
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    • Contributed Indexing:
      Keywords: CRISPR; Protein biochemistry; Protein expression and purification
    • Accession Number:
      0 (Archaeal Proteins)
      0 (Recombinant Proteins)
    • Publication Date:
      Date Created: 20210204 Date Completed: 20220317 Latest Revision: 20220317
    • Publication Date:
      20240829
    • Accession Number:
      PMC7841402
    • Accession Number:
      10.1016/j.xpro.2021.100299
    • Accession Number:
      33537681