title: Crystallization of mutants of Turnip yellow mosaic virus protease/ubiquitin hydrolase designed to prevent protease self-recognition.
abstract: Processing of the polyprotein of Turnip yellow mosaic virus is mediated by the protease PRO. PRO cleaves at two places, one of which is at the C-terminus of the PRO domain of another polyprotein molecule. In addition to this processing activity, PRO possesses an ubiquitin hydrolase (DUB) activity. The crystal structure of PRO has previously been reported in its polyprotein-processing mode with the C-terminus of one PRO inserted into the catalytic site of the next PRO, generating PRO polymers in the crystal packing of the trigonal space group. Here, two mutants designed to disrupt specific PRO-PRO interactions were generated, produced and purified. Crystalline plates were obtained by seeding and cross-seeding from initial `sea urchin'-like microcrystals of one mutant. The plates diffracted to beyond 2 Å resolution at a synchrotron source and complete data sets were collected for the two mutants. Data processing and analysis indicated that both mutant crystals belonged to the same monoclinic space group, with two molecules of PRO in the asymmetric unit.
authors: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris-Sud, 1 Avenue de la Terrasse, 91198 Gif-sur-Yvette, France.
authors: Maya
authors: M
authors: Ayach
authors: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris-Sud, 1 Avenue de la Terrasse, 91198 Gif-sur-Yvette, France.
authors: Stéphane
authors: S
authors: Bressanelli
mesh_terms: D000595:Amino Acid Sequence; D003460:Crystallization; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D017806:Tymovirus / Q000737:chemistry* / Q000201:enzymology* / Q000235:genetics; D025801:Ubiquitin / Q000737:chemistry* / Q000235:genetics; D064570:Ubiquitin-Specific Proteases / Q000737:chemistry* / Q000235:genetics
chemical_list: D025801:Ubiquitin; D006867:Hydrolases; D064570:Ubiquitin-Specific Proteases
keywords: self-processing viral cysteine proteinase; ubiquitin hydrolase