アミドからE・オレフィン対アミドからエステル骨幹のH結合の乱れ:H結合エネルギーを抽出するためのOO排斥を評価する
Yanwen Fu1, Jianmin Gao, Jan Bieschke
1Department of Chemistry and The Skaggs Institute of Chemical Biology, The Scripps Research Institute, BCC 506, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|December 15, 2006
まとめ
研究者は,アミド変異を用いて,タンパク質βシートにおける酸素-酸素排斥を定量化した. この研究では,酸素-酸素の排斥エネルギーを推定し,タンパク質のβシートにおける固有の水素結合の自由エネルギーを計算します.
科学分野:
- タンパク質の構造と折りたたみ
- 生物物理化学 生物物理化学とは
- 分子ダイナミクス 分子ダイナミクス
背景:
- 水素結合は,タンパク質の構造と安定性にとって極めて重要です.
- 酸素-酸素排斥のような特定の相互作用のエネルギー貢献を定量化することは困難です.
- Pin WWドメインは,タンパク質の折り畳みを研究するためのモデルシステムとして機能します.
研究 の 目的:
- ベータシートの文脈で酸素-酸素排斥エネルギー (DeltaGO-Orep) を推定する.
- ピンWW領域における特定の残留物 (F23-R14) の間の固有の水素結合の自由エネルギーを決定する.
- 水素結合を乱すための新しい変異方法を開発し,適用する.
主な方法:
- アミドからエステル,アミドからEオレフィンへの基幹変異戦略を用いる.
- エンジニアリングされたタンパク質ミュータントの熱力学的折りたたみエネルギー測定を用いて.
- 関連するモデル化合物を用いて,移動自由エネルギーの差を計算する.
主要な成果:
- 0.3 kcal/molの推定酸素-酸素排斥エネルギー (DeltaGO-Orep) がベータシートで決定されました.
- 固有のF23-R14水素結合の自由エネルギーが1.3 kcal/molであると計算された.
- 変異方法は,標的の水素結合を効果的に乱した.
結論:
- 酸素-酸素排斥は,タンパク質ベータシートのエネルギー景観に大きく貢献しています.
- タンパク質の固有の水素結合の強さは,これらの生体物理学技術を使用して正確に決定することができます.
- この研究は,タンパク質の折り畳みと安定性を支配する基本的な力についての貴重な洞察を提供します.
関連する概念動画
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Ethers are generally unreactive and unsuitable for direct nucleophilic substitution reactions since the alkoxy groups are strong bases and, therefore, poor leaving groups. However, ethers readily undergo acidic-cleavage reactions. Ethers can be converted to alkyl halides when heated with strong acids such as HBr and HI in a sequence of two substitution reactions.
E2 Reaction: Kinetics and Mechanism
SN2 substitutions and E2 eliminations of alkyl halides proceed via a concerted pathway. While the nucleophile attacks the alpha carbon in SN2 reactions, it functions as a strong base and abstracts a beta hydrogen in the E2 mechanism. The rate-limiting transition state in E2 elimination reactions is characterized by partially broken carbon–hydrogen and carbon–halogen bonds and a partially formed pi bond between the alpha and beta carbons. The beta hydrogen and halide are eliminated...
E2 Reaction: Stereochemistry and Regiochemistry
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When a substrate with two different β hydrogens undergoes an E2 elimination, the presence of a strong base can yield two regioisomeric alkenes. The more-substituted alkene is the major product and...
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Esters to Carboxylic Acids: Acid-Catalyzed Hydrolysis
Hydrolysis of esters under acidic conditions proceeds through a nucleophilic acyl substitution. In the presence of excess water, the reaction proceeds in a reversible manner, forming carboxylic acids and alcohols.
During hydrolysis, the ester is first activated towards nucleophilic attack through the protonation of the carboxyl oxygen atom by the acid catalyst. The protonation makes the ester carbonyl carbon more electrophilic. In the next step, water acts as a nucleophile and adds to the...
During hydrolysis, the ester is first activated towards nucleophilic attack through the protonation of the carboxyl oxygen atom by the acid catalyst. The protonation makes the ester carbonyl carbon more electrophilic. In the next step, water acts as a nucleophile and adds to the...
Reactivity of Enolate Ions
Enolate ions are formed by the acid–base reaction of a carbonyl compound with a base. This leads to deprotonation of the α hydrogen atom, leading to a resonance-stabilized enolate ion where one of the contributing structures is an oxyanion, which imparts additional stability. Therefore, the proton on the α carbon is more acidic in nature than that of other sp3-hybridized C–H bonds but less acidic than those in O–H bonds where the negative charge in the conjugate base is localized on the oxygen...
Amines to Alkenes: Hofmann Elimination
Alkenes can be obtained from amines via an E2 elimination. The amine is first converted into a good leaving group, such as a quaternary ammonium salt. This is accomplished by treating the amine with an excess of alkyl halide, which results in a halide salt. Next, the halide salt is transformed into a hydroxide salt that functions as a base to enable elimination.
Under thermal conditions, the hydroxide can abstract a proton from the β carbon; this generates an alkene with the simultaneous...
Under thermal conditions, the hydroxide can abstract a proton from the β carbon; this generates an alkene with the simultaneous...


