title: Cloning, purification, crystallization and 1.57 Å resolution X-ray data analysis of AmsI, the tyrosine phosphatase controlling amylovoran biosynthesis in the plant pathogen Erwinia amylovora.
abstract: The Gram-negative bacterium Erwinia amylovora is a destructive pathogen of plants belonging to the Rosaceae family. Amongst its pathogenicity factors, E. amylovora produces the exopolysaccharide amylovoran, which contributes to the occlusion of plant vessels, causing wilting of shoots and eventually resulting in plant death. Amylovoran biosynthesis requires the presence of 12 genes (from amsA to amsL) clustered in the ams region of the E. amylovora genome. They mostly encode glycosyl transferases (AmsG, AmsB, AmsD, AmsE, AmsJ and AmsK), proteins involved in amylovoran translocation and assembly (AmsH, AmsL and AmsC), and also a tyrosine kinase (AmsA) and a tyrosine phosphatase (AmsI), which are both involved in the regulation of amylovoran biosynthesis. The low-molecular-weight protein tyrosine phosphatase AmsI was overexpressed as a His6-tagged protein in Escherichia coli, purified and crystallized. X-ray diffraction data were collected to a maximum resolution of 1.57 Å in space group P3121.
authors.affiliation: Laboratory of Bioorganic Chemistry and Bio-Crystallography (B2Cl), Faculty of Science and Technology, Free University of Bolzano, Piazza Università 5, 39100 Bolzano, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Benini
authors.affiliation: Biological Chemistry Department, John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, England.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Caputi
authors.affiliation: EMBL, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Michele
authors.initials: M
authors.lastname: Cianci
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D044084:Erwinia amylovora / Q000378:metabolism*; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis*; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry*
chemical_list: D011135:Polysaccharides, Bacterial; C100769:amylovoran; D017027:Protein Tyrosine Phosphatases
keywords: AmsI; Erwinia; amylovoran; exopolysaccharides; tyrosine phosphatase