Crystal 'unengineering': reducing the crystallisability of sulfolobus solfataricus Hjc

Claire L. Middleton, Joanne L. Parker, Gavin J. Knott, Malcolm F. White, Charles S. Bond (Lead / Corresponding author)

    Research output: Contribution to journalArticlepeer-review

    2 Citations (Scopus)


    The protein Hjc from the thermophilic archaeon Sulfolobus solfataricus (Ss) presented many challenges to both structure solution and formation of stable complexes with its substrate, the DNA four-way or Holliday junction. As the challenges were caused by an uncharacteristically high propensity for rapid and promiscuous crystallisation, we investigated the molecular cause of this behaviour, corrected it by mutagenesis, and solved the X-ray crystal structures of the two mutants. An active site mutant SsHjcA32A crystallised in space group I23 (a 144.2Å; 68% solvent), and a deletion of a key crystal contact site, SsHjcδ62-63 crystallised in space group P21 (a 64.60, b 61.83, c 55.25Å; β≤95.74°; 28% solvent). Characterisation and comparative analysis of the structures are presented along with discussion of the pitfalls of the use of protein engineering to alter crystallisability while maintaining biological function.

    Original languageEnglish
    Pages (from-to)1818-1823
    Number of pages6
    JournalAustralian Journal of Chemistry
    Issue number12
    Publication statusPublished - 18 Aug 2014

    ASJC Scopus subject areas

    • General Chemistry


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