1sm4

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1sm4, resolution 2.50Å

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Crystal Structure Analysis of the Ferredoxin-NADP+ Reductase from Paprika

Overview

cDNA of Capsicum annuum Yolo Wonder (paprika) has been prepared from total cellular RNA, and the complete gene encoding paprika ferredoxin-NADP(+) reductase (pFNR) precursor was sequenced and cloned from this cDNA. Fusion to a T7 promoter allowed expression in Escherichia coli. Both native and recombinant pFNR were purified to homogeneity and crystallized. The crystal structure of pFNR has been solved by Patterson search techniques using the structure of spinach ferredoxin-NADP(+) reductase as search model. The structure was refined at 2.5-A resolution to a crystallographic R-factor of 19.8% (R(free) = 26.5%). The overall structure of pFNR is similar to other members of the ferredoxin-NADP(+) reductase family, the major differences concern a long loop (residues 167-177) that forms part of the FAD binding site and some of the variable loops in surface regions. The different orientation of the FAD binding loop leads to a tighter interaction between pFNR and the adenine moiety of FAD. The physiological redox partners [2Fe-2S]-ferredoxin I and NADP(+) were modeled into the native structure of pFNR. The complexes reveal a protein-protein interaction site that is consistent with existing biochemical data and imply possible orientations for the side chain of tyrosine 362, which has to be displaced by the nicotinamide moiety of NADP(+) upon binding. A reasonable electron transfer pathway could be deduced from the modeled structures of the complexes.

About this Structure

1SM4 is a Single protein structure of sequence from Capsicum annuum with and as ligands. This structure supersedes the now removed PDB entry 1FB3. Active as Ferredoxin--NADP(+) reductase, with EC number 1.18.1.2 Full crystallographic information is available from OCA.

Reference

Crystal structure of paprika ferredoxin-NADP+ reductase. Implications for the electron transfer pathway., Dorowski A, Hofmann A, Steegborn C, Boicu M, Huber R, J Biol Chem. 2001 Mar 23;276(12):9253-63. Epub 2000 Oct 26. PMID:11053431

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