Crystal structures of two tropinone reductases: Different reaction stereospecificities in the same protein fold

被引:114
作者
Nakajima, K [1 ]
Yamashita, A
Akama, H
Nakatsu, T
Kato, H
Hashimoto, T
Oda, J
Yamada, Y
机构
[1] Nara Inst Sci & Technol, Grad Sch Biol Sci, Nara 6300101, Japan
[2] Kyoto Univ, Inst Chem Res, Kyoto 6110011, Japan
关键词
D O I
10.1073/pnas.95.9.4876
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A pair of tropinone reductases (TRs) share 64% of the same amino acid residues and belong to the short-chain dehydrogenase/reductase family. In the synthesis of tropane alkaloids in several medicinal plants, the TRs reduce a carbonyl group of an alkaloid intermediate, tropinone, to hydroxy groups with different diastereomeric configurations. To clarify the structural basis for their different reaction stereospecificities, me determined the crystal structures of the two enzymes at 2.4- and 2.3-Angstrom resolutions. The overall folding of the two enzymes was almost identical. The conservation was not confined within the core domains that are conserved within the protein Family but extended outside the case domain where each family member has its characteristic structure. The binding sites for the cofactor and the positions of the active site residues mere well conserved between the two TRs. The substrate binding site was composed mostly of hydrophobic amino acids in both TRs, but the presence of different charged residues conferred different electrostatic environments on the two enzymes. A modeling study indicated that these charged residues play a major role in controlling the binding orientation of tropinone within the substrate binding site, thereby determining the stereospecificity of the reaction product, The results obtained herein raise the possibility that in certain cases different stereospecificities can be acquired in enzymes by changing a few amino acid residues within substrate binding sites.
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页码:4876 / 4881
页数:6
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