Investigating the role of a backbone to substrate hydrogen bond in OMP decarboxylase using a site-specific amide to ester substitution
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- Bijoy J. Desai
- Departments of aBiochemistry and
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- Yuki Goto
- Department of Chemistry, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan;
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- Alessandro Cembran
- Department of Chemistry, University of Minnesota, Minneapolis, MN 55455;
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- Alexander A. Fedorov
- Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461; and
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- Steven C. Almo
- Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461; and
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- Jiali Gao
- Department of Chemistry, University of Minnesota, Minneapolis, MN 55455;
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- Hiroaki Suga
- Department of Chemistry, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan;
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- John A. Gerlt
- Departments of aBiochemistry and
書誌事項
- 公開日
- 2014-10
- DOI
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- 10.1073/pnas.1411772111
- 公開者
- Proceedings of the National Academy of Sciences
この論文をさがす
説明
<jats:title>Significance</jats:title> <jats:p> Orotidine 5′-monophosphate decarboxylase has attracted intense enzymological interest, because it achieves a very large rate enhancement (∼10 <jats:sup>17</jats:sup> ) without the use of cofactors. Previous studies provided evidence that substrate destabilization and vinyl anion intermediate stabilization contribute to the rate enhancement. Using in vitro translation, we generated a backbone amide to ester substitution to evaluate the importance of the hydrogen bond between a backbone amide and the substrate in intermediate stabilization. The hydrogen bond contributes modestly (≤10 <jats:sup>2</jats:sup> ), suggesting that the intermediate is primarily stabilized by electrostatic interactions with the active site. This study establishes a versatile method for generation of backbone amide to ester substitutions in sufficient quantities to investigate the importance of backbone amide hydrogen bonding interactions in enzyme-catalyzed reactions. </jats:p>
収録刊行物
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- Proceedings of the National Academy of Sciences
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Proceedings of the National Academy of Sciences 111 (42), 15066-15071, 2014-10
Proceedings of the National Academy of Sciences
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キーワード
- Models, Molecular
- Orotidine-5'-Phosphate Decarboxylase
- Static Electricity
- Euryarchaeota
- Molecular Dynamics Simulation
- Crystallography, X-Ray
- Catalysis
- Mass Spectrometry
- Protein Structure, Secondary
- RNA, Transfer
- Tandem Mass Spectrometry
- Catalytic Domain
- Escherichia coli
- RNA, Catalytic
- Cell-Free System
- Temperature
- Esters
- Hydrogen Bonding
- Hydrogen-Ion Concentration
- Amides
- Protein Structure, Tertiary
- Protein Biosynthesis
- Spectrophotometry, Ultraviolet
詳細情報 詳細情報について
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- CRID
- 1362262946224023552
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- ISSN
- 10916490
- 00278424
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- PubMed
- 25275007
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- データソース種別
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- Crossref
- OpenAIRE