Preliminary X-ray characterization of the ribonuclease P (C5 protein) from Escherichia coli: Expression, crystallization and cryoconditions

Hui Woog Choe; Dae Gwin Jeong; Jung Hee Park; Ramona Schlesinger; Jörg Labahn; Klaus Peter Hofmann; Georg Büldt

doi:10.1107/S0907444902020784

Preliminary X-ray characterization of the ribonuclease P (C5 protein) from Escherichia coli: Expression, crystallization and cryoconditions

Hui Woog Choe^*
, Dae Gwin Jeong
, Jung Hee Park
, Ramona Schlesinger
, Jörg Labahn
, Klaus Peter Hofmann
, Georg Büldt

^*Corresponding author for this work

Bioresource and Biomolecule Engineering

Jeonbuk National University
Humboldt University of Berlin
Inst. Biol. Informationsverarbeitung

Research output: Contribution to journal › Journal article › peer-review

4 Scopus citations

Abstract

The gene for Escherichia coli ribonuclease P (RNase P) protein (also known as C5 protein) and its mutant C5-C113A have been expressed as GST fusion proteins in E. coli at a high level. After cleavage of the fusion protein, highly purified functional C5 protein is obtained that can be crystallized with 2.5-2.6 M (NH₄)₂HPO₄/(NH₄)H₂PO ₄ pH 7.0 at room temperature. These crystals are suitable for X-ray analysis, belong to the space group P3₁21 or P3₂21 (unit-cell parameters a = b = 66.67, c = 142.09 Å) and diffract to 2.9 Å at 100 K using sorbitol and glycerol as cryoprotectants. For three molecules in the asymmetric unit a V_M of 2.17 Å³ Da^-1 was calculated.

Original language	English
Pages (from-to)	350-352
Number of pages	3
Journal	Acta Crystallographica - Section D Biological Crystallography
Volume	59
Issue number	2
DOIs	https://doi.org/10.1107/S0907444902020784
State	Published - 2003.02.1

Quacquarelli Symonds(QS) Subject Topics

Biological Sciences

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@article{6e391088ca114e52a3a74deeefab8b7a,

title = "Preliminary X-ray characterization of the ribonuclease P (C5 protein) from Escherichia coli: Expression, crystallization and cryoconditions",

abstract = "The gene for Escherichia coli ribonuclease P (RNase P) protein (also known as C5 protein) and its mutant C5-C113A have been expressed as GST fusion proteins in E. coli at a high level. After cleavage of the fusion protein, highly purified functional C5 protein is obtained that can be crystallized with 2.5-2.6 M (NH4)2HPO4/(NH4)H2PO 4 pH 7.0 at room temperature. These crystals are suitable for X-ray analysis, belong to the space group P3121 or P3221 (unit-cell parameters a = b = 66.67, c = 142.09 {\AA}) and diffract to 2.9 {\AA} at 100 K using sorbitol and glycerol as cryoprotectants. For three molecules in the asymmetric unit a VM of 2.17 {\AA}3 Da-1 was calculated.",

author = "Choe, \{Hui Woog\} and Jeong, \{Dae Gwin\} and Park, \{Jung Hee\} and Ramona Schlesinger and J{\"o}rg Labahn and Hofmann, \{Klaus Peter\} and Georg B{\"u}ldt",

year = "2003",

month = feb,

day = "1",

doi = "10.1107/S0907444902020784",

language = "English",

volume = "59",

pages = "350--352",

journal = "Acta Crystallographica - Section D Biological Crystallography",

issn = "0907-4449",

number = "2",

}

Preliminary X-ray characterization of the ribonuclease P (C5 protein) from Escherichia coli: Expression, crystallization and cryoconditions. / Choe, Hui Woog; Jeong, Dae Gwin; Park, Jung Hee et al.
In: Acta Crystallographica - Section D Biological Crystallography, Vol. 59, No. 2, 01.02.2003, p. 350-352.

Research output: Contribution to journal › Journal article › peer-review

TY - JOUR

T1 - Preliminary X-ray characterization of the ribonuclease P (C5 protein) from Escherichia coli

T2 - Expression, crystallization and cryoconditions

AU - Choe, Hui Woog

AU - Jeong, Dae Gwin

AU - Park, Jung Hee

AU - Schlesinger, Ramona

AU - Labahn, Jörg

AU - Hofmann, Klaus Peter

AU - Büldt, Georg

PY - 2003/2/1

Y1 - 2003/2/1

N2 - The gene for Escherichia coli ribonuclease P (RNase P) protein (also known as C5 protein) and its mutant C5-C113A have been expressed as GST fusion proteins in E. coli at a high level. After cleavage of the fusion protein, highly purified functional C5 protein is obtained that can be crystallized with 2.5-2.6 M (NH4)2HPO4/(NH4)H2PO 4 pH 7.0 at room temperature. These crystals are suitable for X-ray analysis, belong to the space group P3121 or P3221 (unit-cell parameters a = b = 66.67, c = 142.09 Å) and diffract to 2.9 Å at 100 K using sorbitol and glycerol as cryoprotectants. For three molecules in the asymmetric unit a VM of 2.17 Å3 Da-1 was calculated.

AB - The gene for Escherichia coli ribonuclease P (RNase P) protein (also known as C5 protein) and its mutant C5-C113A have been expressed as GST fusion proteins in E. coli at a high level. After cleavage of the fusion protein, highly purified functional C5 protein is obtained that can be crystallized with 2.5-2.6 M (NH4)2HPO4/(NH4)H2PO 4 pH 7.0 at room temperature. These crystals are suitable for X-ray analysis, belong to the space group P3121 or P3221 (unit-cell parameters a = b = 66.67, c = 142.09 Å) and diffract to 2.9 Å at 100 K using sorbitol and glycerol as cryoprotectants. For three molecules in the asymmetric unit a VM of 2.17 Å3 Da-1 was calculated.

UR - https://www.scopus.com/pages/publications/0037321931

U2 - 10.1107/S0907444902020784

DO - 10.1107/S0907444902020784

M3 - Journal article

C2 - 12554950

AN - SCOPUS:0037321931

SN - 0907-4449

VL - 59

SP - 350

EP - 352

JO - Acta Crystallographica - Section D Biological Crystallography

JF - Acta Crystallographica - Section D Biological Crystallography

IS - 2

ER -

Preliminary X-ray characterization of the ribonuclease P (C5 protein) from Escherichia coli: Expression, crystallization and cryoconditions

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