Tags

Type your tag names separated by a space and hit enter

Purification and crystallization of Escherichia coli oligoribonuclease.
Acta Crystallogr D Biol Crystallogr. 2004 Apr; 60(Pt 4):736-9.AC

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 A resolution and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 70.43, b = 72.87, c = 147.76 A, 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 (P2(1)2(1)2(1); a = 63.74, b = 74.31, c = 74.19 A). Finally, a hexagonal crystal form was obtained using sodium acetate as a precipitant (a = 91.5, b = 91.5, c = 111.1 A). This crystal (Zn-ApUp-Orn) belongs to the P6(5) space group and has three oligoribonuclease molecules per asymmetric unit.

Authors+Show Affiliations

Fiedler TJ
Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, PO Box 016129, Miami, FL 33101-6129, USA.
Vincent HA
No affiliation info available
Zuo Y
No affiliation info available
Gavrialov O
No affiliation info available
Malhotra A
No affiliation info available

MeSH

Cloning, MolecularCrystallizationCrystallography, X-RayDimerizationEscherichia coli ProteinsExoribonucleasesManganese

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15039570

Citation

Fiedler, Tristan J., et al. "Purification and Crystallization of Escherichia Coli Oligoribonuclease." Acta Crystallographica. Section D, Biological Crystallography, vol. 60, no. Pt 4, 2004, pp. 736-9.
Fiedler TJ, Vincent HA, Zuo Y, et al. Purification and crystallization of Escherichia coli oligoribonuclease. Acta Crystallogr D Biol Crystallogr. 2004;60(Pt 4):736-9.
Fiedler, T. J., Vincent, H. A., Zuo, Y., Gavrialov, O., & Malhotra, A. (2004). Purification and crystallization of Escherichia coli oligoribonuclease. Acta Crystallographica. Section D, Biological Crystallography, 60(Pt 4), 736-9.
Fiedler TJ, et al. Purification and Crystallization of Escherichia Coli Oligoribonuclease. Acta Crystallogr D Biol Crystallogr. 2004;60(Pt 4):736-9. PubMed PMID: 15039570.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Purification and crystallization of Escherichia coli oligoribonuclease. AU - Fiedler,Tristan J, AU - Vincent,Helen A, AU - Zuo,Yuhong, AU - Gavrialov,Orit, AU - Malhotra,Arun, Y1 - 2004/03/23/ PY - 2003/02/07/received PY - 2004/01/27/accepted PY - 2004/3/25/pubmed PY - 2004/12/16/medline PY - 2004/3/25/entrez SP - 736 EP - 9 JF - Acta crystallographica. Section D, Biological crystallography JO - Acta Crystallogr D Biol Crystallogr VL - 60 IS - Pt 4 N2 - 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 A resolution and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 70.43, b = 72.87, c = 147.76 A, 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 (P2(1)2(1)2(1); a = 63.74, b = 74.31, c = 74.19 A). Finally, a hexagonal crystal form was obtained using sodium acetate as a precipitant (a = 91.5, b = 91.5, c = 111.1 A). This crystal (Zn-ApUp-Orn) belongs to the P6(5) space group and has three oligoribonuclease molecules per asymmetric unit. SN - 0907-4449 UR - https://www.unboundmedicine.com/medline/citation/15039570/Purification_and_crystallization_of_Escherichia_coli_oligoribonuclease_ DB - PRIME DP - Unbound Medicine ER -