The non-structural protein Nsp15 from the aetiological agent of SARS (severe acute respiratory syndrome) has recently been characterized as a uridine-specific endoribonuclease. This enzyme plays an essential role in viral replication and transcription since a mutation in the related H229E human coronavirus nsp15 gene can abolish viral RNA synthesis. SARS full-length Nsp15 (346 amino acids) has been cloned and expressed in Escherichia coli with an N-terminal hexahistidine tag and has been purified to homogeneity. The protein was subsequently crystallized using PEG 8000 or 10 000 as precipitants. Small cubic crystals of the apoenzyme were obtained from 100 nl nanodrops. They belong to space group P4132 or P4332, with unit-cell parameters a = b = c = 166.8 Å. Diffraction data were collected to a maximum resolution of 2.7 Å.

Crystallization and preliminary X-ray diffraction analysis of Nsp15 from SARS coronavirus / S. Ricagno, B. Coutard, S. Grisel, N. Brémond, K. Dalle, F. Tocque, V. Campanacci, J. Lichière, V. Lantez, C. Debarnot, C. Cambillau, B. Canard, M. Egloff. - In: ACTA CRYSTALLOGRAPHICA. SECTION F, STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS. - ISSN 1744-3091. - 62:4(2006 Apr), pp. 409-411.

Crystallization and preliminary X-ray diffraction analysis of Nsp15 from SARS coronavirus

S. Ricagno
Primo
;
2006

Abstract

The non-structural protein Nsp15 from the aetiological agent of SARS (severe acute respiratory syndrome) has recently been characterized as a uridine-specific endoribonuclease. This enzyme plays an essential role in viral replication and transcription since a mutation in the related H229E human coronavirus nsp15 gene can abolish viral RNA synthesis. SARS full-length Nsp15 (346 amino acids) has been cloned and expressed in Escherichia coli with an N-terminal hexahistidine tag and has been purified to homogeneity. The protein was subsequently crystallized using PEG 8000 or 10 000 as precipitants. Small cubic crystals of the apoenzyme were obtained from 100 nl nanodrops. They belong to space group P4132 or P4332, with unit-cell parameters a = b = c = 166.8 Å. Diffraction data were collected to a maximum resolution of 2.7 Å.
cloning, molecular; crystallization; escherichia coli; macromolecular substances; peptide fragments; protein conformation; rna replicase; recombinant proteins; sars virus; viral nonstructural proteins; x-ray diffraction; structural biology; biophysics; biochemistry; genetics; condensed matter physics
Settore BIO/10 - Biochimica
apr-2006
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/492688
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