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High-resolution studies of hydride transfer in the ferredoxin:NADP+ reductase superfamily.
FEBS J. 2017 10; 284(19):3302-3319.FJ

Abstract

Ferredoxin: NADP+ reductase (FNR) is an FAD-containing enzyme best known for catalysing the transfer of electrons from ferredoxin (Fd) to NADP+ to make NADPH during photosynthesis. It is also the prototype for a broad enzyme superfamily, including the NADPH oxidases (NOXs) that all catalyse similar FAD-enabled electron transfers between NAD(P)H and one-electron carriers. Here, we define further mechanistic details of the NAD(P)H ⇌ FAD hydride-transfer step of the reaction based on spectroscopic studies and high-resolution (~ 1.5 Å) crystallographic views of the nicotinamide-flavin interaction in crystals of corn root FNR Tyr316Ser and Tyr316Ala variants soaked with either nicotinamide, NADP+ , or NADPH. The spectra obtained from FNR crystal complexes match those seen in solution and the complexes reveal active site packing interactions and patterns of covalent distortion of the FAD that imply significant active site compression that would favour catalysis. Furthermore, anisotropic B-factors show that the mobility of the C4 atom of the nicotinamide in the FNR:NADP+ complex has a directionality matching that expected for boat-like excursions of the nicotinamide ring thought to enhance hydride transfer. Arguments are made for the relevance of this binding mode to catalysis, and specific consideration is given to how the results extrapolate to provide insight to structure-function relations for the membrane-bound NOX enzymes for which little structural information has been available.

DATABASES

Structural data are available in the PDB database under the accession numbers 3LO8 (wild-type), 5VW4 [Y316S:nicotinamide (P32 21)], 5VW9 [Y316S:nicotinamide (P31 21)], 5VW3 [Y316S:NADP+ (P32 21)], 5VW8 [Y316S:NADP+ (P31 21)], 5VW2 [Y316S:NADPH (P32 21)], 5VW5 [Y316A:nicotinamide (P32 21)], 5VW6 [Y316A:NADP+ (P32 21)], 5VW7 [Y316A:NADPH (P32 21)], 5VWA [Y316F (P32 21)], and 5VWB [Y316F:NADP+ (P31 21)]. Enzyme Commission number: ferredoxin:NADP+ reductase - E C1.18.1.2.

Authors+Show Affiliations

Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.Department of Biosciences, Università degli Studi di Milano, Italy.Department of Biosciences, Università degli Studi di Milano, Italy.Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.Department of Biosciences, Università degli Studi di Milano, Italy.Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

28783258

Citation

Kean, Kelsey M., et al. "High-resolution Studies of Hydride Transfer in the ferredoxin:NADP+ Reductase Superfamily." The FEBS Journal, vol. 284, no. 19, 2017, pp. 3302-3319.
Kean KM, Carpenter RA, Pandini V, et al. High-resolution studies of hydride transfer in the ferredoxin:NADP+ reductase superfamily. FEBS J. 2017;284(19):3302-3319.
Kean, K. M., Carpenter, R. A., Pandini, V., Zanetti, G., Hall, A. R., Faber, R., Aliverti, A., & Karplus, P. A. (2017). High-resolution studies of hydride transfer in the ferredoxin:NADP+ reductase superfamily. The FEBS Journal, 284(19), 3302-3319. https://doi.org/10.1111/febs.14190
Kean KM, et al. High-resolution Studies of Hydride Transfer in the ferredoxin:NADP+ Reductase Superfamily. FEBS J. 2017;284(19):3302-3319. PubMed PMID: 28783258.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - High-resolution studies of hydride transfer in the ferredoxin:NADP+ reductase superfamily. AU - Kean,Kelsey M, AU - Carpenter,Russell A, AU - Pandini,Vittorio, AU - Zanetti,Giuliana, AU - Hall,Andrea R, AU - Faber,Rick, AU - Aliverti,Alessandro, AU - Karplus,P Andrew, Y1 - 2017/08/29/ PY - 2017/06/12/received PY - 2017/07/28/revised PY - 2017/08/03/accepted PY - 2017/8/8/pubmed PY - 2017/10/21/medline PY - 2017/8/8/entrez KW - NADPH oxidase KW - enzyme mechanism KW - flavoenzyme KW - hydride transfer KW - protein crystallography SP - 3302 EP - 3319 JF - The FEBS journal JO - FEBS J VL - 284 IS - 19 N2 - : Ferredoxin: NADP+ reductase (FNR) is an FAD-containing enzyme best known for catalysing the transfer of electrons from ferredoxin (Fd) to NADP+ to make NADPH during photosynthesis. It is also the prototype for a broad enzyme superfamily, including the NADPH oxidases (NOXs) that all catalyse similar FAD-enabled electron transfers between NAD(P)H and one-electron carriers. Here, we define further mechanistic details of the NAD(P)H ⇌ FAD hydride-transfer step of the reaction based on spectroscopic studies and high-resolution (~ 1.5 Å) crystallographic views of the nicotinamide-flavin interaction in crystals of corn root FNR Tyr316Ser and Tyr316Ala variants soaked with either nicotinamide, NADP+ , or NADPH. The spectra obtained from FNR crystal complexes match those seen in solution and the complexes reveal active site packing interactions and patterns of covalent distortion of the FAD that imply significant active site compression that would favour catalysis. Furthermore, anisotropic B-factors show that the mobility of the C4 atom of the nicotinamide in the FNR:NADP+ complex has a directionality matching that expected for boat-like excursions of the nicotinamide ring thought to enhance hydride transfer. Arguments are made for the relevance of this binding mode to catalysis, and specific consideration is given to how the results extrapolate to provide insight to structure-function relations for the membrane-bound NOX enzymes for which little structural information has been available. DATABASES: Structural data are available in the PDB database under the accession numbers 3LO8 (wild-type), 5VW4 [Y316S:nicotinamide (P32 21)], 5VW9 [Y316S:nicotinamide (P31 21)], 5VW3 [Y316S:NADP+ (P32 21)], 5VW8 [Y316S:NADP+ (P31 21)], 5VW2 [Y316S:NADPH (P32 21)], 5VW5 [Y316A:nicotinamide (P32 21)], 5VW6 [Y316A:NADP+ (P32 21)], 5VW7 [Y316A:NADPH (P32 21)], 5VWA [Y316F (P32 21)], and 5VWB [Y316F:NADP+ (P31 21)]. Enzyme Commission number: ferredoxin:NADP+ reductase - E C1.18.1.2. SN - 1742-4658 UR - https://www.unboundmedicine.com/medline/citation/28783258/High_resolution_studies_of_hydride_transfer_in_the_ferredoxin:NADP+_reductase_superfamily_ L2 - https://doi.org/10.1111/febs.14190 DB - PRIME DP - Unbound Medicine ER -