サイト固有のプラチナ (((II) リボ酵素活性部位におけるクロスリンク
Erich G Chapman1, Victoria J DeRose
1Department of Chemistry and Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403, USA.
Journal of the American Chemical Society
|November 11, 2011
まとめ
研究者らは,酸ヒオ酸代用を用いた新しいプラチナ (II) RNA-RNAクロスリンク方法を開発した. このテクニックは,Hammerhead ribozymeで実証されたように,RNAのサイト固有の構造分析を可能にします.
科学分野:
- バイオケミストリー バイオケミストリー
- 分子生物学は分子生物学である.
- 構造生物学 構造生物学とは
背景:
- RNAの構造とダイナミクスは,機能にとって極めて重要です.
- サイト固有のクロスリンクは,RNA構造生物学にとって貴重な距離制約を提供します.
- 既存の方法は,正確なRNA構造の決定のためにさらに精細化する必要があります.
研究 の 目的:
- プラチナ (II) 複合体を用いた新しいRNA-RNAクロスリンク戦略を導入する.
- ハンマーヘッドリボ酵素の構造を調査するこの方法の適用を調査する.
- サイト固有のRNAの構造と動的研究のための新しいツールを確立する.
主な方法:
- 新しいRNA-RNAクロスリンク戦略は,Pt(II) ターゲティングによるフォスフォロチオアート置換をベースにしています.
- シス-ダイアミン Pt (II) 複合体をリン酸塩酸塩置換RNAに動態的に誘導し,固定する.
- Pt(II) 複合置換によるRNA核塩基間の金属媒介クロスリンクを誘導する.
主要な成果:
- Pt(II) クロスリンクは,ハマーヘッドリボエンザイム活性部位内のG8およびG10核酸で,リン酸塩酸がスチーシール結合にあるときに発生しました.
- 30%の収量に達するクロスリンクの効率は,Mg2+) イオン濃度とRNAの三次構造形成に依存していた.
- 交叉リンクは,RNAヘリクではなく,活性部位内で特に観察され,部位特異性を示しました.
結論:
- 開発されたPt (II) ベースのクロスリンク戦略は,サイト固有のRNA構造分析に有効です.
- この方法は,RNAの構造と動態,特に複雑なRNAモチーフの探索のための有望な新しいツールを提供します.
- この発見は,RNAの構造生物学を進歩させるための,運動的に惰性であるPt (II) 複合体の可能性を強調している.
関連する概念動画
Ribozymes
The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Ribozymes can be...
Ribozymes
The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Ribozymes can be...
Riboswitches
Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
Ligand Binding and Linkage
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 the...
Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...


