Crystallization and preliminary X-ray diffraction studies of Hje, a Holliday junction resolving enzyme from Sulfolobus solfataricus

CL Middleton, JL Parker, DJ Richard, Malcolm Frederick White, CS Bond

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

Holliday junction endonuclease (Hje) from Sulfolobus solfataricus is a resolving enzyme involved in cleaving specific sites on either side of recombinant four-way Holliday junctions. The HJE gene from S. solfataricus was cloned from genomic DNA into the pET19b Escherichia coli expression vector and recombinant protein was expressed to high levels. Hje was purified using heat treatment, cation exchange and gel filtration. Hanging-drop crystallization trials yielded primitive hexagonal crystals which diffract to 2.4 Angstrom on a laboratory source. Systematic absences (only 00l = 6n present) and poor scaling in P622 indicate that the space group is P6(1) or its enantiomer. Failed attempts at molecular replacement using models of a related archaeal resolving enzyme, Hjc, raise the possibility of a difference in quaternary structure between Hjc and Hje, which may be responsible for differences in their activities.

Original languageEnglish
Pages (from-to)171-173
Number of pages3
JournalActa Crystallographica. Section D, Biological crystallography
Volume59 (Part 1)
DOIs
Publication statusPublished - Jan 2003

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