Aldose and aldehyde reductases: structure-function studies on the coenzyme and inhibitor-binding sites.
OBJECTIVE
To identify the structural features responsible for the differences in coenzyme and inhibitor specificities of aldose and aldehyde reductases.


METHODS
The crystal structure of porcine aldehyde reductase in complex with NADPH and the aldose reductase inhibitor sorbinil was determined. The contribution of each amino acid lining the coenzyme-binding site to the binding of NADPH was calculated using the Discover package. In human aldose reductase, the role of the non-conserved Pro 216 (Ser in aldehyde reductase) in the binding of coenzyme was examined by site-directed mutagenesis.


RESULTS
Sorbinil binds to the active site of aldehyde reductase and is hydrogen-bonded to Trp 22, Tyr 50, His 113, and the non-conserved Arg 312. Unlike tolrestat, the binding of sorbinil does not induce a change in the side chain conformation of Arg 312. Mutation of Pro 216 to Ser in aldose reductase makes the binding of coenzyme more similar to that of aldehyde reductase.


CONCLUSIONS
The participation of non-conserved active site residues in the binding of inhibitors and the differences in the structural changes required for the binding to occur are responsible for the differences in the potency of inhibition of aldose and aldehyde reductases. We report that the non-conserved Pro 216 in aldose reductase contributes to the tight binding of NADPH.
El-Kabbani
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Department of Medicinal Chemistry, Victorian College of Pharmacy, Monash University, Parkville, Australia. ossama.el-kabbani@vcp.monash.edu.au
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Ginell
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D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D003067:Coenzymes / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D006801:Humans; D007093:Imidazoles / Q000737:chemistry / Q000378:metabolism; D048289:Imidazolidines; D016297:Mutagenesis, Site-Directed; D009249:NADP / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013552:Swine
D003067:Coenzymes; D007093:Imidazoles; D048289:Imidazolidines; D009249:NADP; D000449:Aldehyde Reductase; C026411:sorbinil