プロテインの選択的金属サイト誘導アリレーション
Jens Willwacher1, Ritu Raj1, Shabaz Mohammed1
1Department of Chemistry, Chemistry Research Laboratory, University of Oxford , Mansfield Road, Oxford OX1 3TA, U.K.
Journal of the American Chemical Society
|June 24, 2016
まとめ
この研究は,金属結合タンパク質とその環境の識別を可能にする,サイト選択性タンパク質改変のためのパラジアム触媒反応を導入する. この方法は,金属依存の酵素活性を理解するのに役立ちます.
科学分野:
- 化学生物学
- プロテオミクス
- カタリシス
背景:
- 金属結合タンパク質は様々な生物学的プロセスで 重要な役割を果たします
- タンパク質の機能を明らかにするために,金属結合部位の特定の環境を理解することは不可欠です.
- 金属結合部位とその周辺を特定するための現在の方法は,範囲と適用が制限されている可能性があります.
研究 の 目的:
- タンパク質の部位選択的改変のための新しいパラジアム媒介S-アリレーション法を開発する.
- 自然な金属結合モチーフを活用して 反応性残留物を正確に標的にします
- 金属結合タンパク質の化学的識別と結合部位環境の特徴づけを可能にする.
主な方法:
- パラジウム触媒によるS-アリレーション反応を利用した.
- タンパク質内の固有金属結合モチーフを直接アリレーションに利用した.
- タンパク質のアリレーション部位を特定するためにプロテオミック分析を用いた.
- 細胞溶解体にこの方法を適用し,酵素調節におけるその有用性を調査した.
主要な成果:
- 金属結合部位に近接するアリレーションの高いサイト選択性を達成した.
- 金属を結合するタンパク質を成功裏に特定し,これらの場所の微小環境を特徴づけた.
- 標準条件下でのS-アリレーション変換の広範囲と容易さを実証した.
- 細胞溶解物のような複雑な生物学的マトリックスでの適用性を示した.
結論:
- 開発されたパラジアム媒介のS-アリレーションは,化学生物学とプロテオミクスの強力なツールである.
- この方法は,タンパク質における金属結合部位の識別と特徴付けを容易にする.
- このアプローチは,金属依存酵素とその調節を研究するための多角的な戦略を提供します.
さらに関連する動画
関連する概念動画
00:49Ligand Binding and Linkage
5.9K
Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...
5.9K
02:40Ligand Binding Sites
15.7K
Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
15.7K
01:19Allosteric Proteins-ATCase
6.9K
Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
6.9K
02:17Reduction of Alkenes: Asymmetric Catalytic Hydrogenation
4.0K
Catalytic hydrogenation of alkenes is a transition-metal catalyzed reduction of the double bond using molecular hydrogen to give alkanes. The mode of hydrogen addition follows syn stereochemistry.
The metal catalyst used can be either heterogeneous or homogeneous. When hydrogenation of an alkene generates a chiral center, a pair of enantiomeric products is expected to form. However, an enantiomeric excess of one of the products can be facilitated using an enantioselective reaction or an...
The metal catalyst used can be either heterogeneous or homogeneous. When hydrogenation of an alkene generates a chiral center, a pair of enantiomeric products is expected to form. However, an enantiomeric excess of one of the products can be facilitated using an enantioselective reaction or an...
4.0K
01:22Preparation of 1° Amines: Azide Synthesis
4.8K
Direct alkylation of ammonia produces polyalkylated amines, along with a quaternary ammonium salt. To exclusively prepare primary amines, the azide synthesis method can be used.
Azide ions act as good nucleophiles and react with unhindered alkyl halides to form alkyl azides. Alkyl azides do not participate in further nucleophilic substitution reactions, thereby eliminating the chances of polyalkylated products. Alkyl azides are reduced by hydride-based reducing agents, like lithium aluminum...
Azide ions act as good nucleophiles and react with unhindered alkyl halides to form alkyl azides. Alkyl azides do not participate in further nucleophilic substitution reactions, thereby eliminating the chances of polyalkylated products. Alkyl azides are reduced by hydride-based reducing agents, like lithium aluminum...
4.8K
01:30Preparation of Amines: Alkylation of Ammonia and Amines
4.9K
Alkylation is one of the methods used to prepare amines. Direct alkylation of ammonia or a primary amine with an alkyl halide gives polyalkylated amines along with a quaternary ammonium salt through successive SN2 reactions. This process of making the quaternary salt through the direct alkylation method is called exhaustive alkylation.
Each alkylation step makes the nitrogen center more nucleophilic, which triggers successive alkylations until a quaternary ammonium salt is formed. Considering...
Each alkylation step makes the nitrogen center more nucleophilic, which triggers successive alkylations until a quaternary ammonium salt is formed. Considering...
4.9K