title: Expression, purification, crystallization and preliminary X-ray crystallographic analysis of a major fragment of the resuscitation-promoting factor RpfB from Mycobacterium tuberculosis.
abstract: RpfB is required for the virulence and the resuscitation from dormancy of Mycobacterium tuberculosis, the bacterium responsible for tuberculosis. This protein is a cell-wall glycosidase that acts by cleaving peptidoglycans of the bacterial cell wall and therefore stimulates both bacterial growth and resuscitation from latency. RpfB consists of 362 residues organized into five domains. A long portion of RpfB, including its C-terminal catalytic domain, the G5 domain and one of its three DUF348 domains, which are of hitherto unknown structure and function, has been successfully crystallized using vapour-diffusion methods and seeding techniques. The crystals diffracted to 2.55 Å resolution and belonged to space group C222(1), with unit-cell parameters a=102.3, b=126.2, c=85.87 Å. Model building using phases derived from the combined use of multiwavelength anomalous dispersion and molecular replacement is in progress. The results obtained here will provide the first structural characterization of a DUF348 domain reported to date and will shed light on the functional role of the noncatalytic domains of RpfB.
authors.affiliation: Institute of Biostructures and Bioimaging, CNR, Via Mezzocannone 16, I-80134 Napoli, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Ruggiero
authors.forename: Flavia
authors.initials: F
authors.lastname: Squeglia
authors.forename: Luciano
authors.initials: L
authors.lastname: Pirone
authors.forename: Stefania
authors.initials: S
authors.lastname: Correale
authors.forename: Rita
authors.initials: R
authors.lastname: Berisio
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D016207:Cytokines / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation
chemical_list: D001426:Bacterial Proteins; D016207:Cytokines; C113704:resuscitation-promoting factor, bacteria