Crystal structure of the flavoenzyme PA4991 from Pseudomonas aeruginosa.
Acta Crystallogr F Struct Biol Commun
; 72(Pt 2): 105-11, 2016 Feb.
Article
em En
| MEDLINE
| ID: mdl-26841760
ABSTRACT
The locus PA4991 in Pseudomonas aeruginosa encodes an open reading frame that has been identified as essential for the virulence and/or survival of this pathogenic organism in the infected host. Here, it is shown that this gene encodes a monomeric FAD-binding protein of molecular mass 42.2â
kDa. The structure of PA4991 was determined by a combination of molecular replacement using a search model generated with Rosetta and phase improvement by a low-occupancy heavy-metal derivative. PA4991 belongs to the GR2 family of FAD-dependent oxidoreductases, comprising an FAD-binding domain typical of the glutathione reductase family and a second domain dominated by an eight-stranded mixed ß-sheet. Most of the protein-FAD interactions are via the FAD-binding domain, but the isoalloxazine ring is located at the domain interface and interacts with residues from both domains. A comparison with the structurally related glycine oxidase and glycerol-3-phosphate dehydrogenase shows that in spite of very low amino-acid sequence identity (<18%) several active-site residues involved in substrate binding in these enzymes are conserved in PA4991. However, enzymatic assays show that PA4991 does not display amino-acid oxidase or glycerol-3-phosphate dehydrogenase activities, suggesting that it requires different substrates for activity.
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MEDLINE
Assunto principal:
Pseudomonas aeruginosa
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Cristalografia por Raios X
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Cristalização
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Flavoproteínas
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article