Crystal structures of Leptospira interrogans FAD-containing ferredoxin-NADP+ reductase and its complex with NADP+

Nascimento, Alessandro S.; Catalano-Dupuy, Daniela L.; Bernardes, Amanda; Neto, Mário de Oliveira; Santos, M. A. M.; Ceccarelli, Eduardo A.; Polikarpov, Igor

doi:10.1186/1472-6807-7-69

Cited by 20 publications

(31 citation statements)

References 36 publications

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“…We have previously assigned the FNR from Leptospira interrogans (LepFNR) to the plastidic FNR class [2], and this was confirmed by resolution of the LepFNR crystal structure [9]. The presence of a plastidic FNR in Leptospira might be explained as the result of lateral gene transfer, and this was probably selected to provide the bacterium with some adaptive advantages.…”

Section: Introductionmentioning

confidence: 82%

A Highly Stable Plastidic-Type Ferredoxin-NADP(H) Reductase in the Pathogenic Bacterium Leptospira interrogans

et al. 2011

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Leptospira interrogans is a bacterium that is capable of infecting animals and humans, and its infection causes leptospirosis with a range of symptoms from flu-like to severe illness and death. Despite being a bacteria, Leptospira interrogans contains a plastidic class ferredoxin-NADP(H) reductase (FNR) with high catalytic efficiency, at difference from the bacterial class FNRs. These flavoenzymes catalyze the electron transfer between NADP(H) and ferredoxins or flavodoxins. The inclusion of a plastidic FNR in Leptospira metabolism and in its parasitic life cycle is not currently understood. Bioinformatic analyses of the available genomic and proteins sequences showed that the presence of this enzyme in nonphotosynthetic bacteria is restricted to the Leptospira genus and that a [4Fe-4S] ferredoxin (LB107) encoded by the Leptospira genome may be the natural substrate of the enzyme. Leptospira FNR (LepFNR) displayed high diaphorase activity using artificial acceptors and functioned as a ferric reductase. LepFNR displayed cytochrome c reductase activity with the Leptospira LB107 ferredoxin with an optimum at pH 6.5. Structural stability analysis demonstrates that LepFNR is one of the most stable FNRs analyzed to date. The persistence of a native folded LepFNR structure was detected in up to 6 M urea, a condition in which the enzyme retains 38% activity. In silico analysis indicates that the high LepFNR stability might be due to robust interactions between the FAD and the NADP+ domains of the protein. The limited bacterial distribution of plastidic class FNRs and the biochemical and structural properties of LepFNR emphasize the uniqueness of this enzyme in the Leptospira metabolism. Our studies show that in L. interrogans a plastidic-type FNR exchanges electrons with a bacterial-type ferredoxin, process which has not been previously observed in nature.

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Section: Introductionmentioning

confidence: 82%

A Highly Stable Plastidic-Type Ferredoxin-NADP(H) Reductase in the Pathogenic Bacterium Leptospira interrogans

et al. 2011

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“…Besides, Rubisco is a well-studied enzyme because of its extensive agricultural and environmental significance (Selesi et al, 2005). Ferredoxin-NADP(H) oxidoreductase (F84) can catalyze the electron transfer between NADP(H) and the proteins ferredoxin or flavodoxin (Thomas et al, 2006;Nascimento et al, 2007). Phosphoribulokinase (F105) is found in most phototrophic organisms.…”

Section: Blastxmentioning

confidence: 99%

Expression of special genes inhibited by powdery mildew (Blumeria graminis f. sp. tritici) in wheat germplasm N9436

et al. 2010

Front. Agric. China

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Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important fungal diseases in common wheat (Triticum aestivum L.) worldwide. Wheat germplasm N9436 is resistant to powdery mildew. In the present study, a backward subtracted cDNA library was constructed with cDNA from N9436 leaves inoculated by Blumeria graminis as the driver and cDNA from uninoculated N9436 leaves as the tester. A total of 120 positive clones were randomly chosen from the SSHcDNA library and were amplified with sp6 and t7 primers to examine the insert size. After screening the repeated and redundant sequences, 59 expressed sequence tags (EST) were acquired. Nucleic acid and protein homology search were performed using the basic local alignment search tool (BLAST) program with the default settings at NCBI website (http://www.ncbi.nlm.nih.gov). BlastX results in nr-protein database revealed that 23 ESTs were highly homologous with known proteins involved in primary metabolism, energy metabolism, transport, signal transduction, and disease resistance and defenses. BlastNr results showed that 47 and 10 ESTs had high identities with known Unigene and function-unknown ESTs, respectively, and two ESTs matched none in the nrdatabase. Twenty-one ESTs were both in the nucleic acid and protein databases, including seven ESTs associated with powdery mildew resistance. Among them, one was responsible for signal transduction and six for systemic acquired resistance (SAR) system.

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“…Enzyme kinetics were monitored by using an Optizen 2120 UV/VIS spectrophotometer (Mecasys, Korea) at 25°C under anaerobic conditions. The ferric reductase assay was (13,28). Each kinetic experiment was repeated five times.…”

Section: Methodsmentioning

confidence: 99%

Ferredoxin-NADP ⁺ Reductase from Pseudomonas putida Functions as a Ferric Reductase

et al. 2009

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Pseudomonas putida harbors two ferredoxin-NADP؉ reductases (Fprs) on its chromosome, and their functions remain largely unknown. Ferric reductase is structurally contained within the Fpr superfamily. Interestingly, ferric reductase is not annotated on the chromosome of P. putida. In an effort to elucidate the function of the Fpr as a ferric reductase, we used a variety of biochemical and physiological methods using the wild-type and mutant strains. In both the ferric reductase and flavin reductase assays, FprA and FprB preferentially used NADPH and NADH as electron donors, respectively. Two Fprs prefer a native ferric chelator to a synthetic ferric chelator and utilize free flavin mononucleotide (FMN) as an electron carrier. FprB has a higher k cat /K m value for reducing the ferric complex with free FMN. The growth rate of the fprB mutant was reduced more profoundly than that of the fprA mutant, the growth rate of which is also lower than the wild type in ferric iron-containing minimal media. Flavin reductase activity was diminished completely when the cell extracts of the fprB mutant plus NADH were utilized, but not the fprA mutant with NADPH. This indicates that other NADPH-dependent flavin reductases may exist. Interestingly, the structure of the NAD(P) region of FprB, but not of FprA, resembled the ferric reductase (Fre) of Escherichia coli in the homology modeling. This study demonstrates, for the first time, the functions of Fprs in P. putida as flavin and ferric reductases. Furthermore, our results indicated that FprB may perform a crucial role as a NADHdependent ferric/flavin reductase under iron stress conditions.

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Crystal structures of Leptospira interrogans FAD-containing ferredoxin-NADP+ reductase and its complex with NADP+

Cited by 20 publications

References 36 publications

A Highly Stable Plastidic-Type Ferredoxin-NADP(H) Reductase in the Pathogenic Bacterium Leptospira interrogans

A Highly Stable Plastidic-Type Ferredoxin-NADP(H) Reductase in the Pathogenic Bacterium Leptospira interrogans

Expression of special genes inhibited by powdery mildew (Blumeria graminis f. sp. tritici) in wheat germplasm N9436

Ferredoxin-NADP ⁺ Reductase from Pseudomonas putida Functions as a Ferric Reductase

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Crystal structures of Leptospira interrogans FAD-containing ferredoxin-NADP+ reductase and its complex with NADP+

Cited by 20 publications

References 36 publications

A Highly Stable Plastidic-Type Ferredoxin-NADP(H) Reductase in the Pathogenic Bacterium Leptospira interrogans

A Highly Stable Plastidic-Type Ferredoxin-NADP(H) Reductase in the Pathogenic Bacterium Leptospira interrogans

Expression of special genes inhibited by powdery mildew (Blumeria graminis f. sp. tritici) in wheat germplasm N9436

Ferredoxin-NADP + Reductase from Pseudomonas putida Functions as a Ferric Reductase

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Ferredoxin-NADP ⁺ Reductase from Pseudomonas putida Functions as a Ferric Reductase