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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
Author(s) -
Benini Stefano,
Caputi Lorenzo,
Cianci Michele
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14024947
Subject(s) - erwinia , biology , pathogen , protein tyrosine phosphatase , microbiology and biotechnology , phosphatase , biochemistry , escherichia coli , tyrosine , biosynthesis , botany , gene , phosphorylation
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 ams A to ams L) 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 His 6 ‐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 P 3 1 21.