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Expression, purification and crystallization of the SARS-CoV macro domain.


ABSTRACT: 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 P2(1), with unit-cell parameters a = 37.5, b = 55.6, c = 108.9 angstroms, beta = 91.4 degrees, and the asymmetric unit contains either two or three molecules. Both native and selenomethionine-labelled crystals diffract to 1.8 angstroms.

SUBMITTER: Malet H 

PROVIDER: S-EPMC2222557 | biostudies-literature | 2006 Apr

REPOSITORIES: biostudies-literature

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Expression, purification and crystallization of the SARS-CoV macro domain.

Malet Hélène H   Dalle Karen K   Brémond Nicolas N   Tocque Fabienne F   Blangy Stéphanie S   Campanacci Valérie V   Coutard Bruno B   Grisel Sacha S   Lichière Julie J   Lantez Violaine V   Cambillau Christian C   Canard Bruno B   Egloff Marie Pierre MP  

Acta crystallographica. Section F, Structural biology and crystallization communications 20060325 Pt 4


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) e  ...[more]

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