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Title: The purification, crystallization and preliminary structural characterization of FAD-dependent monooxygenase PhzS, a phenazine-modifying enzyme from Pseudomonas aeruginosa

Author
item GOHAIN, NEELAKSHI - ASDASDFASDF
item Thomashow, Linda
item MAVRODI, D - WASHINGTON STATE UNIV.
item BLANKENFELDT, W - MAX PLANCK INST.

Submitted to: Acta Crystallographica Section B: Structural Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/20/2006
Publication Date: 10/1/2006
Citation: Gohain, N., Thomashow, L.S., Mavrodi, D.V., Blankenfeldt, W. 2006. The purification, crystallization and preliminary structural characterization of FAD-dependent monooxygenase PhzS, a phenazine-modifying enzyme from Pseudomonas aeruginosa. Acta Crystallographica F 62:989-992

Interpretive Summary: The blue chloroform-soluble bacterial metabolite pyocyanin contributes to the survival and virulence of Pseudomonas aeruginosa, an important pathogen of burn victims and immunocompromised people. Little is known about two enzymes, designated PhzM and PhzS, that function in the synthesis of pyocyanin. In this study, the PhzS enzyme was purified and crystallized. Native crystals contained two copies of the enzyme in each enzymatic unit. The study provides valuable new information about how this enzyme functions in pyocyanin biosynthesis.

Technical Abstract: The blue chloroform-soluble bacterial metabolite pyocyanin (1-hydroxy-5-methyl-phenazine) contributes to the survival and virulence of Pseudomonas aeruginosa, an important Gram-negative opportunistic pathogen of humans and animals. Little is known about the two enzymes, designated PhzM and PhzS, that function in the synthesis of pyocyanin from phenazine-1-carboxylic acid. In this study, the FAD-dependent monooxygenase PhzS was purified and crystallized from lithium sulfate/ammonium sulfate/sodium citrate pH 5.5. Native crystals belong to space group C2, with unit-cell parameters a = 144.2, b= 96.2, c= 71.7 angstrom, alpha = gamma = 90, beta = 110.5 degrees. They contain two monomers of PhzS in the asymmetric unit and diffract to a resolution of 2.4 angstrom. Seleno-L-methionine-labelled PhzS also crystallizes in space group C2, but the unit-cell parameters change to a = 70.6, b = 76.2, c = 80.2 angstrom, alpha = gamma = 90, beta = 110.5 degrees and the diffraction limit is 2.7 angstrom.