Open AccessExpression, purification and crystallization of the SARS‐CoV macro domain
Author(s) -
Malet Hélène,
Dalle Karen,
Brémond Nicolas,
Tocque Fabienne,
Blangy Stéphanie,
Campanacci Valérie,
Coutard Bruno,
Grisel Sacha,
Lichière Julie,
Lantez Violaine,
Cambillau Christian,
Canard Bruno,
Egloff MariePierre
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106009274
Subject(s) - escherichia coli , crystallization , biology , protein domain , gene , crystallography , coronavirus , chemistry , computational biology , biochemistry , covid-19 , organic chemistry , medicine , disease , pathology , infectious disease (medical specialty)
Macro domains or X domains are found as modules of multidomain proteins, but can also constitute a protein on their own. Recently, biochemical and structural studies of cellular macro domains have been performed, showing that they are active as ADP‐ribose‐1′′‐phosphatases. Macro domains are also present in a number of positive‐stranded RNA viruses, but their precise function in viral replication is still unknown. The major human pathogen severe acute respiratory syndrome coronavirus (SARS‐CoV) encodes 16 non‐structural proteins (nsps), one of which (nsp3) encompasses a macro domain. The SARS‐CoV nsp3 gene region corresponding to amino acids 182–355 has been cloned, expressed in Escherichia coli , purified and crystallized. The crystals belong to space group P 2 1 , with unit‐cell parameters a = 37.5, b = 55.6, c = 108.9 Å, β = 91.4°, and the asymmetric unit contains either two or three molecules. Both native and selenomethionine‐labelled crystals diffract to 1.8 Å.