Structure determination of an FMN reductase from Pseudomonas aeruginosa PA01 using sulfur anomalous signal

Rakhi Agarwal, Jeffrey B. Bonanno, Stephen K. Burley, Subramanyam Swaminathan

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

The availability of high-intensity synchrotron facilities, technological advances in data-collection techniques and improved data-reduction and crystallographic software have ushered in a new era in high-throughput macromolecular crystallography. Here, the de novo automated crystal structure determination at 1.28 Å resolution of an NAD(P)H-dependent FMN reductase flavoprotein from Pseudomonas aeruginosa PA01-derived protein Q9I4D4 using the anomalous signal from an unusually small number of S atoms is reported. Although this protein lacks the flavodoxin key fingerprint motif [(T/S)XTGXT], it has been confirmed to bind flavin mononucleotide and the binding site was identified via X-ray crystallography. This protein contains a novel flavin mononucleotide-binding site GSLRSGSYN, which has not been previously reported. Detailed statistics pertaining to sulfur phasing and other factors contributing to structure determination are discussed. Structural comparisons of the apoenzyme and the protein complexed with flavin mononucleotide show conformational changes on cofactor binding. NADPH-dependent activity has been confirmed with biochemical assays.

Original languageEnglish (US)
Pages (from-to)383-391
Number of pages9
JournalActa Crystallographica Section D: Biological Crystallography
Volume62
Issue number4
DOIs
StatePublished - Apr 1 2006

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ASJC Scopus subject areas

  • Structural Biology

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