title: Purification, crystallization and preliminary X-ray diffraction analysis of GatD, a glutamine amidotransferase-like protein from Staphylococcus aureus peptidoglycan.
abstract: Amidation of peptidoglycan is an essential feature in Staphylococcus aureus that is necessary for resistance to β-lactams and lysozyme. GatD, a 27 kDa type I glutamine amidotransferase-like protein, together with MurT ligase, catalyses the amidation reaction of the glutamic acid residues of the peptidoglycan of S. aureus. The native and the selenomethionine-derivative proteins were crystallized using the sitting-drop vapour-diffusion method with polyethylene glycol, sodium acetate and calcium acetate. The crystals obtained diffracted beyond 1.85 and 2.25 Å, respectively, and belonged to space group P212121. X-ray diffraction data sets were collected at Diamond Light Source (on beamlines I02 and I04) and were used to obtain initial phases.
authors.affiliation: Macromolecular Crystallography Group and GlycoLab, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal.
authors.forename: Diana
authors.initials: D
authors.lastname: Vieira
authors.affiliation: Laboratory of Molecular Genetics, Microbiology of Human Pathogens Unit, Instituto de Tecnologia Química e Biológica da Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Teresa A
authors.initials: TA
authors.lastname: Figueiredo
authors.affiliation: Oxford Protein Production Facility, Research Complex at Harwell, Didcot, England.
authors.forename: Anil
authors.initials: A
authors.lastname: Verma
authors.affiliation: Laboratory of Molecular Genetics, Microbiology of Human Pathogens Unit, Instituto de Tecnologia Química e Biológica da Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Rita G
authors.initials: RG
authors.lastname: Sobral
authors.affiliation: Laboratory of Molecular Genetics, Microbiology of Human Pathogens Unit, Instituto de Tecnologia Química e Biológica da Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Ludovice
authors.affiliation: Laboratory of Molecular Genetics, Microbiology of Human Pathogens Unit, Instituto de Tecnologia Química e Biológica da Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Hermínia
authors.initials: H
authors.lastname: de Lencastre
authors.affiliation: Diamond Light Source, Didcot, England.
authors.forename: Jose
authors.initials: J
authors.lastname: Trincao
mesh_terms: D000595:Amino Acid Sequence; D003460:Crystallization; D008969:Molecular Sequence Data; D010457:Peptidoglycan / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics; D000637:Transaminases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014961:X-Ray Diffraction
chemical_list: D010457:Peptidoglycan; D000637:Transaminases; C020656:glutamine-pyruvate aminotransferase
keywords: GatD; Staphylococcus aureus; glutamine amidotransferase-like protein