Purification and crystallization of Escherichia coli oligoribonuclease

Tristan J. Fiedler, Helen A. Vincent, Yuhong Zuo, Orit Gavrialov, Arun Malhotra

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Oligoribonuclease (Orn) is an essential 3′-to-5′ hydrolytic exoribonuclease which degrades short oligoribonucleotides to 5′ mononucleotides. Escherichia coli Orn has been crystallized under several different conditions using ammonium sulfate, sodium citrate and sodium acetate as precipitants. Both native and selenomethionine-labeled oligoribonuclease (SeMet-Orn) can be crystallized at room temperature in 1.4-1.55 M sodium citrate. The SeMet-Orn crystals diffract to 2.2 Å resolution and belong to space group P212121, with unit-cell parameters a = 70.43, b = 72.87, c = 147.76 Å, and two dimers in the asymmetric unit. When grown in the presence of manganese, a second crystal form (Mn-SeMet-Orn) was obtained containing a single dimer per asymmetric unit (P212121; a = 63.74, b = 74.31, c = 74.19 Å). Finally, a hexagonal crystal form was obtained using sodium acetate as a precipitant (a = 91.5, b = 91.5, c = 111.1 Å). This crystal (Zn-ApUp-Orn) belongs to the P65 space group and has three oligoribonuclease molecules per asymmetric unit.

Original languageEnglish
Pages (from-to)736-739
Number of pages4
JournalActa Crystallographica Section D: Biological Crystallography
Volume60
Issue number4
DOIs
StatePublished - Apr 1 2004

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Escherichia
Crystallization
purification
Escherichia coli
Purification
sodium
crystallization
Selenomethionine
citrates
crystals
acetates
dimers
Sodium Acetate
Crystals
ammonium sulfates
Dimers
manganese
Oligoribonucleotides
Ammonium Sulfate
Manganese

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics
  • Condensed Matter Physics
  • Structural Biology

Cite this

Purification and crystallization of Escherichia coli oligoribonuclease. / Fiedler, Tristan J.; Vincent, Helen A.; Zuo, Yuhong; Gavrialov, Orit; Malhotra, Arun.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 60, No. 4, 01.04.2004, p. 736-739.

Research output: Contribution to journalArticle

Fiedler, Tristan J. ; Vincent, Helen A. ; Zuo, Yuhong ; Gavrialov, Orit ; Malhotra, Arun. / Purification and crystallization of Escherichia coli oligoribonuclease. In: Acta Crystallographica Section D: Biological Crystallography. 2004 ; Vol. 60, No. 4. pp. 736-739.
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