4KOA

Crystal Structure Analysis of 1,5-anhydro-D-fructose reductase from Sinorhizobium meliloti

Summary for 4KOA

• Entry DOI: 10.2210/pdb4koa/pdb
• Related: 2GLX
• Descriptor: 1,5-anhydro-D-fructose reductase, NADPH DIHYDRO-NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE (3 entities in total)
• Functional Keywords: gfo/idh/moca family, dehydrogenase, sugar binding, oxidoreductase
• Biological source: Sinorhizobium meliloti (Ensifer meliloti)
• Total number of polymer chains: 1
• Total formula weight: 35638.89

Authors

Schu, M.,Faust, A.,Stosik, B.,Kohring, G.-W.,Giffhorn, F.,Scheidig, A.J. (deposition date: 2013-05-11, release date: 2013-08-07, Last modification date: 2023-09-20)

Primary citation

Schu, M.,Faust, A.,Stosik, B.,Kohring, G.W.,Giffhorn, F.,Scheidig, A.J.
The structure of substrate-free 1,5-anhydro-D-fructose reductase from Sinorhizobium meliloti 1021 reveals an open enzyme conformation.
Acta Crystallogr.,Sect.F, 69:844-849, 2013

PubMed Abstract: 1,5-Anhydro-D-fructose (1,5-AF) is an interesting building block for enantioselective and stereoselective organic synthesis. Enzymes acting on this compound are potential targets for structure-based protein/enzyme design to extend the repertoire of catalytic modifications of this and related building blocks. Recombinant 1,5-anhydro-D-fructose reductase (AFR) from Sinorhizobium meliloti 1021 was produced in Escherichia coli, purified using a fused 6×His affinity tag and crystallized in complex with the cofactor NADP(H) using the hanging-drop technique. Its structure was determined to 1.93 Å resolution using molecular replacement. The structure displays an empty substrate-binding site and can be interpreted as an open conformation reflecting the enzyme state shortly after the release of product, presumably with bound oxidized cofactor NADP⁺. Docking simulations indicated that amino-acid residues Lys94, His151, Trp162, Arg163, Asp176 and His180 are involved in substrate binding, catalysis or product release. The side chain of Lys94 seems to have the ability to function as a molecular switch. The crystal structure helps to characterize the interface relevant for dimer formation as observed in solution. The crystal structure is compared with the structure of the homologue from S. morelense, which was solved in a closed conformation and for which dimer formation in solution could not be verified but seems to be likely based on the presented studies of S. meliloti AFR.

PubMed: 23908025
DOI: 10.1107/S1744309113019490

Experimental method

X-RAY DIFFRACTION (1.93 Å)