Expression, high-pressure refolding, purification, crystallization and preliminary X-ray analysis of a novel single-strand-specific 3′-5′ exonuclease PhoExo I from Pyrococcus horikoshii OT3

Ken Ichi Miyazono, Kanae Tsutsumi, Yoshizumi Ishino, Masaru Tanokura

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

PhoExo I is a single-strand-specific 3′-5′ exonuclease from Pyrococcus horikoshii OT3 and is thought to be involved in a Thermococcales-specific DNA-repair pathway. The recombinant PhoExo I protein was produced as inclusion bodies in Escherichia coli cells. Solubilization of the inclusion bodies was performed by the high-pressure refolding method and highly purified protein was subjected to crystallization by the sitting-drop vapour-diffusion method at 20°C. A crystal of PhoExo I was obtained in a reservoir solution consisting of 0.1 M Tris-HCl pH 8.9, 27% PEG 6000 and diffracted X-rays to 1.52 Å resolution. The crystal of PhoExo I belonged to space group H32, with unit-cell parameters a = b = 112.07, c = 202.28 Å. The crystal contained two PhoExo I molecules in the asymmetric unit.

Original languageEnglish
Pages (from-to)1076-1079
Number of pages4
JournalActa Crystallographica Section F:Structural Biology Communications
Volume70
Issue number8
DOIs
Publication statusPublished - Aug 2014

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

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