2X3U

Ferredoxin-NADP reductase mutant with Tyr 303 replaced by Phe (Y303F)

Summary for 2X3U

• Entry DOI: 10.2210/pdb2x3u/pdb
• Related: 1B2R 1BJK 1BQE 1E62 1E63 1E64 1EWY 1GJR 1GO2 1GR1 1H42 1H85 1OGI 1OGJ 1QGY 1QGZ 1QH0 1QUE 1QUF 1W34 1W35 1W87 2BMW 2BSA 2VYQ 2VZL
• Descriptor: FERREDOXIN-NADP REDUCTASE, FLAVIN-ADENINE DINUCLEOTIDE, SULFATE ION, ... (5 entities in total)
• Functional Keywords: oxidoreductase, flavoprotein
• Biological source: ANABAENA SP.
• Cellular location: Cellular thylakoid membrane; Peripheral membrane protein; Cytoplasmic side: P21890
• Total number of polymer chains: 1
• Total formula weight: 35000.37

Authors

Herguedas, B.,Martinez-Julvez, M.,Hermoso, J.A.,Peregrina, J.R.,Sanchez-Azqueta, A.,Medina, M. (deposition date: 2010-01-27, release date: 2010-05-26, Last modification date: 2023-12-20)

Primary citation

Peregrina, J.R.,Sanchez-Azqueta, A.,Herguedas, B.,Martinez-Julvez, M.,Medina, M.
Role of Specific Residues in Coenzyme Binding, Charge-Transfer Complex Formation, and Catalysis in Anabaena Ferredoxin Nadp(+)-Reductase.
Biochim.Biophys.Acta, 1797:1638-, 2010

PubMed Abstract: Two transient charge-transfer complexes (CTC) form prior and upon hydride transfer (HT) in the reversible reaction of the FAD-dependent ferredoxin-NADP+ reductase (FNR) with NADP+/H, FNR(ox)-NADPH (CTC-1), and FNR(rd)-NADP+ (CTC-2). Spectral properties of both CTCs, as well as the corresponding interconversion HT rates, are here reported for several Anabaena FNR site-directed mutants. The need for an adequate initial interaction between the 2'P-AMP portion of NADP+/H and FNR that provides subsequent conformational changes leading to CTC formation is further confirmed. Stronger interactions between the isoalloxazine and nicotinamide rings might relate with faster HT processes, but exceptions are found upon distortion of the active centre. Thus, within the analyzed FNR variants, there is no strict correlation between the stability of the transient CTCs formation and the rate of the subsequent HT. Kinetic isotope effects suggest that, while in the WT, vibrational enhanced modulation of the active site contributes to the tunnel probability of HT; complexes of some of the active site mutants with the coenzyme hardly allow the relative movement of isoalloxazine and nicotinamide rings along the HT reaction. The architecture of the WT FNR active site precisely contributes to reduce the stacking probability between the isoalloxazine and nicotinamide rings in the catalytically competent complex, modulating the angle and distance between the N5 of the FAD isoalloxazine and the C4 of the coenzyme nicotinamide to values that ensure efficient HT processes.

PubMed: 20471952
DOI: 10.1016/J.BBABIO.2010.05.006

Experimental method

X-RAY DIFFRACTION (1.93 Å)