Mass Spectrometry on Hydrogen/Deuterium Exchange of Escherichia coli Dihydrofolate Reductase: Effects of Loop Mutations
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- Yamamoto Tatsuya
- Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University
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- Izumi Shunsuke
- Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University
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- Ohmae Eiji
- Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University
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- Gekko Kunihiko
- Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University
書誌事項
- タイトル別名
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- Mass Spectrometry of Hydrogen/Deuterium Exchange of <i>Escherichia coli</i> Dihydrofolate Reductase: Effects of Loop Mutations
- Mass Spectrometry of Hydrogen/Deuterium Exchange of Escherichia coli Dihydrofolate Reductase: Effects of Loop Mutations
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説明
To address the effects of single amino acid substitutions on the structural fluctuation of Escherichia coli dihydrofolate reductase (DHFR), hydrogen/deuterium exchange kinetics were investigated at 15°C with wild-type and mutant DHFRs at Gly 67 (six mutants) and Gly 121 (eight mutants) located in two flexible loops, by means of electrospray ionization mass spectrometry. These mutations induced significant changes in the first-order rate constant of proton exchange, kex (0.10-0.27min-1), the number of fast-exchangeable protons, ΔMo (164-222 Da), and the number of protons protected from exchange, ΔM∞ (15-56 Da), relative to the corresponding values for the wild-type enzyme (kex=0.18min-1, ΔMo=164 Da, and ΔM∞=50.5 Da). These kinetic parameters were strongly correlated with the volume of introduced amino acids, but partly correlated with adiabatic compressibility (volume fluctuation), stability, and enzymatic activity. These results indicate that the local structure change due to a single amino acid substitution in loop regions is dramatically magnified to affect the structural fluctuation of the whole DHFR molecule, resulting in complicated changes in its stability and function.
収録刊行物
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- The Journal of Biochemistry
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The Journal of Biochemistry 135 (4), 487-494, 2004
社団法人 日本生化学会
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詳細情報 詳細情報について
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- CRID
- 1390001204966395392
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- NII論文ID
- 130003534687
- 10016199363
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- NII書誌ID
- AA00694073
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- COI
- 1:CAS:528:DC%2BD2cXltFCgtL8%3D
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- ISSN
- 17562651
- 0021924X
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- NDL書誌ID
- 6931153
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- PubMed
- 15115773
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- 抄録ライセンスフラグ
- 使用不可