Structure of rat aldose reductase-like protein AKR1B14 holoenzyme: Probing the role of His269 in coenzyme binding by site-directed mutagenesis
✍ Scribed by Krithika Sundaram; Urmi Dhagat; Satoshi Endo; Roland Chung; Toshiyuki Matsunaga; Akira Hara; Ossama El-Kabbani
- Publisher
- Elsevier Science
- Year
- 2011
- Tongue
- English
- Weight
- 353 KB
- Volume
- 21
- Category
- Article
- ISSN
- 0960-894X
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✦ Synopsis
Rat aldose reductase-like protein (AKR1B14) is the ortholog of mouse vas deferens protein (AKR1B7) playing roles in detoxification of reactive aldehydes and synthesis of prostaglandin F 2a . The crystal structure of the binary complex (AKR1B14-NADPH) was determined at 1.86 Å resolution, and showed that the adenine ring and the 2 0 -phosphate group of the coenzyme formed p-stacking and electrostatic interactions with the imidazole ring and ND1 atom, respectively, of His269, which is not conserved in other aldose reductase-like proteins. The interactions were supported by site-directed mutagenesis of His269 to Arg, Phe and Met, which increased the K m for NADPH by 4, 7 and 127-fold, respectively. This is the first report of the tertiary structure of a rodent AKR1B7 ortholog, which describes the role of a novel dual interaction for the non-conserved His269 in coenzyme binding.