Journal Article

PreJuSER-26893

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Preliminary X-ray characterization of the ribonuclease P (C5 protein) from Escherichia coli : expression, crystallization and cryoconditions

Choe, B. J. ; Jeong, D.-G. ; Park, K. S. ; Schlesinger, R.FZJ* ; Labahn, J.FZJ* ; Hofmann, K. P. ; Büldt, G.FZJ*

2003
Munksgaard Copenhagen

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Please use a persistent id in citations: doi:10.1107/S0907444902020784

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 (NH(4))(2)HPO(4)/(NH(4))H(2)PO(4) pH 7.0 at room temperature. These crystals are suitable for X-ray analysis, belong to the space group P3(1)21 or P3(2)21 (unit-cell parameters a = b = 66.67, c = 142.09 A) and diffract to 2.9 A at 100 K using sorbitol and glycerol as cryoprotectants. For three molecules in the asymmetric unit a V(M) of 2.17 A(3) Da(-1) was calculated.

Keyword(s): Amino Acid Substitution (MeSH) ; Cryoprotective Agents: chemistry (MeSH) ; Crystallization (MeSH) ; Crystallography, X-Ray (MeSH) ; Endoribonucleases: chemistry (MeSH) ; Endoribonucleases: genetics (MeSH) ; Endoribonucleases: metabolism (MeSH) ; Escherichia coli: enzymology (MeSH) ; Escherichia coli Proteins (MeSH) ; Glutathione Transferase: genetics (MeSH) ; Glycerol: chemistry (MeSH) ; RNA, Catalytic: chemistry (MeSH) ; RNA, Catalytic: genetics (MeSH) ; RNA, Catalytic: metabolism (MeSH) ; Recombinant Fusion Proteins: biosynthesis (MeSH) ; Recombinant Fusion Proteins: genetics (MeSH) ; Ribonuclease P (MeSH) ; Ribonucleoproteins: chemistry (MeSH) ; Ribonucleoproteins: genetics (MeSH) ; Ribonucleoproteins: metabolism (MeSH) ; Sorbitol: chemistry (MeSH) ; Cryoprotective Agents ; Escherichia coli Proteins ; RNA, Catalytic ; Recombinant Fusion Proteins ; Ribonucleoproteins ; Sorbitol ; Glycerol ; Glutathione Transferase ; Endoribonucleases ; Ribonuclease P ; ribonuclease P, E coli ; J

Note: Record converted from VDB: 12.11.2012

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Contributing Institute(s):

1. Biologische Strukturforschung (IBI-2)

Research Program(s):

1. Neurowissenschaften (L01)

Appears in the scientific report 2003

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