Ferredoxin/ferredoxin–thioredoxin reductase complex: Complete NMR mapping of the interaction site on ferredoxin by gallium substitution

Xu, Xiaozheng; Kim, Sung-Kun; Schürmann, Peter; Hirasawa, Masakazu; Tripathy, Jatindra N.; Smith, J.P.; Knaff, David B.; Ubbink, Marcellus

doi:10.1016/j.febslet.2006.11.027

Cited by 25 publications

(21 citation statements)

References 36 publications

(42 reference statements)

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“…Recent NMR chemical shift mapping studies of the Fdx-FTR interaction (303) are consistent with the previous results obtained by differential chemical modification (57) and also with the crystal structure of the Fdx-FTR complex (50). The results emphasize the dependency of the interaction on polar as well as nonpolar residues, the latter forming a hydrophobic core.…”

Section: Fig 3 Multiple Sequence Alignment Of the Catalytic Subunitsupporting

confidence: 79%

“…This conclusion was supported by mutagenesis experiments in which the replacement of this C-terminal glutamate residue resulted in a protein incapable of reducing FTR (129). These observations have been corroborated and extended by the recent structural studies (50,303) discussed below. Work during the past several years has also added a new dimension to the function of Fdx.…”

Section: A Ferredoxinsupporting

confidence: 71%

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The Ferredoxin/Thioredoxin System of Oxygenic Photosynthesis

Schürmann

Buchanan

2008

Antioxidants & Redox Signaling

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390

368

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Section: Fig 3 Multiple Sequence Alignment Of the Catalytic Subunitsupporting

confidence: 79%

Section: A Ferredoxinsupporting

confidence: 71%

The Ferredoxin/Thioredoxin System of Oxygenic Photosynthesis

Schürmann

Buchanan

2008

Antioxidants & Redox Signaling

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390

368

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“…Interactions between PetF and various redox partners usually involve three polypeptide motifs (28) -Glu 123 in PetF of C. reinhardtii. These three motifs contain highly conserved acidic residues that direct the first stages of complex generation dominated by electrostatic attraction and intermolecular salt bridge formation (22,29,30).…”

mentioning

confidence: 99%

Characterization of the Key Step for Light-driven Hydrogen Evolution in Green Algae

Winkler

Kuhlgert

Hippler

et al. 2009

Journal of Biological Chemistry

115

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Under anaerobic conditions, several species of green algae perform a light-dependent hydrogen production catalyzed by a special group of [FeFe] hydrogenases termed HydA. Although highly interesting for biotechnological applications, the direct connection between photosynthetic electron transport and hydrogenase activity is still a matter of speculation. By establishing an in vitro reconstitution system, we demonstrate that the photosynthetic ferredoxin (PetF) is essential for efficient electron transfer between photosystem I and HydA1. To investigate the electrostatic interaction process and electron transfer between PetF and HydA1, we performed site-directed mutagenesis. Kinetic analyses with several site-directed mutagenesis variants of HydA1 and PetF enabled us to localize the respective contact sites. These experiments in combination with in silico docking analyses indicate that electrostatic interactions between the conserved HydA1 residue Lys 396 and the C terminus of PetF as well as between the PetF residue Glu 122 and the N-terminal amino group of HydA1 play a major role in complex formation and electron transfer. Mapping of relevant HydA1 and PetF residues constitutes an important basis for manipulating the physiological photosynthetic electron flow in favor of light-driven H 2 production.Among all photosynthetic organisms, only green algae can couple light-driven electron transport originating from water splitting with hydrogen production (1). Hydrogen evolution in the unicellular green alga Chlamydomonas reinhardtii is naturally induced upon nutrient deprivation (2). Especially in the absence of sulfur, the photosynthetic oxygen evolution rate drops below the respiratory rate leading to intracellular anaerobiosis. Under anaerobic conditions, the oxygen-sensitive (2, 3) [FeFe] hydrogenase HydA is synthesized and catalyzes light-dependent H 2 production, thereby dissipating excess redox equivalents under conditions in which the Calvin cycle is down-regulated (4).The extraordinarily small monomeric [FeFe] hydrogenases of green algae only consist of the catalytic core unit containing the active site (H-cluster), whereas other [FeFe] hydrogenases possess an additional N-terminal F-domain harboring one to four accessory iron-sulfur clusters (5, 6). Because Chlorophytatype [FeFe] hydrogenases lack any accessory clusters (7, 8), a direct electron transfer between the native electron donor and the H-cluster has been assumed (9, 10). In C. reinhardtii, HydA1 has been shown to be localized in the chloroplast stroma (11), and first kinetic examinations with purified proteins demonstrated that the plastidic ferredoxin PetF can interact with HydA1. These results and the fact that H 2 production in C. reinhardtii is photosystem I (PSI) 2 -dependent (4) led to the hypothesis that PetF is the native electron donor of the plastidic hydrogenase (12). Derived from in silico analyses, two possible PetF-HydA2 electron transfer complex models were recently suggested (13). However, in contrast to the well studied interacti...

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“…Our recent NMR study identified the bindingi nterface of PetF with HydA1 from C. reinhardtii. [15] Apart from the study of the interaction with ferredoxin thioredoxin reductase, [14] none of theseN MR spectroscopic investigations provided information about the important residues coordinating and surrounding the [2 Fe-2 S] cluster.B ecause of the proximity of these residues to the cluster, their nuclear spinsr elax quickly,a nd their NMR signals become broad or undetectable. This paramagnetic relaxation enhancement (PRE) can be avoided by substitution of the paramagnetic iron with diamagnetic gallium(III).…”

mentioning

confidence: 99%

“…(FNR), [13] and ferredoxin thioredoxin reductase [14] has been identified by NMR-titration experiments. Our recent NMR study identified the bindingi nterface of PetF with HydA1 from C. reinhardtii.…”

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confidence: 99%

Structural Insight into the Complex of Ferredoxin and [FeFe] Hydrogenase from Chlamydomonas reinhardtii

et al. 2015

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The transfer of photosynthetic electrons by the ferredoxin PetF to the [FeFe] hydrogenase HydA1 in the microalga Chlamydomonas reinhardtii is a key step in hydrogen production. Electron delivery requires a specific interaction between PetF and HydA1. However, because of the transient nature of the electron-transfer complex, a crystal structure remains elusive. Therefore, we performed protein-protein docking based on new experimental data from a solution NMR spectroscopy investigation of native and gallium-substituted PetF. This provides valuable information about residues crucial for complex formation and electron transfer. The derived complex model might help to pinpoint residue substitution targets for improved hydrogen production.

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Ferredoxin/ferredoxin–thioredoxin reductase complex: Complete NMR mapping of the interaction site on ferredoxin by gallium substitution

Cited by 25 publications

References 36 publications

The Ferredoxin/Thioredoxin System of Oxygenic Photosynthesis

The Ferredoxin/Thioredoxin System of Oxygenic Photosynthesis

Characterization of the Key Step for Light-driven Hydrogen Evolution in Green Algae

Structural Insight into the Complex of Ferredoxin and [FeFe] Hydrogenase from Chlamydomonas reinhardtii

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