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関連する概念動画

¹H NMR: Complex Splitting01:13

¹H NMR: Complex Splitting

1.7K
A proton M that is coupled to a proton X results in doublet signals for M. However, NMR-active nuclei can be simultaneously coupled to more than one nonequivalent nucleus. When M is coupled to a second proton A, such as in styrene oxide, each peak in the doublet is split into another doublet.
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied...
1.7K
¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)01:20

¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)

1.3K
When proton-coupled carbon-13 spectra are simplified by a broadband proton decoupling technique, structural information about the coupled protons is lost. Distortionless enhancement by polarization transfer (DEPT) is a technique that provides information on the number of hydrogens attached to each carbon in a molecule. While the DEPT experiment utilizes complex pulse sequences, the pulse delay and flip angle are specifically manipulated. The resulting signals have different phases depending on...
1.3K
¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR01:15

¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR

1.4K
The axial and equatorial protons in cyclohexane can be distinguished by performing a variable-temperature NMR experiment. In this process, except for one proton, the remaining eleven protons are replaced by deuterium. The deuterium substitution avoids the possible peak splitting caused by the spin-spin coupling between the adjacent protons. The remaining proton flips between the axial and equatorial positions.
1.4K
¹³C NMR: ¹H–¹³C Decoupling01:04

¹³C NMR: ¹H–¹³C Decoupling

1.7K
The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...
1.7K
Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

1.5K
In an NMR sample, precise measurement of the absolute absorption frequencies of nuclei is difficult. A standard internal reference compound is added, and the frequency difference between the reference signal and sample signals is measured.
The internal reference compound generally used in NMR spectroscopy is tetramethylsilane (TMS). TMS is preferred because it is chemically inert, soluble in NMR solvents, and easily removable. Also, the highly shielded methyl protons in TMS yield an intense...
1.5K
Double Resonance Techniques: Overview01:12

Double Resonance Techniques: Overview

870
Double resonance techniques in Nuclear Magnetic Resonance (NMR) spectroscopy involve the simultaneous application of two different frequencies or radiofrequency pulses to manipulate and observe two distinct nuclear spins. One important application of double resonance is spin decoupling, which selectively suppresses coupling with one type of nucleus while observing the NMR signal from another nucleus, simplifying the spectrum and enhancing resolution.
Spin decoupling is usually achieved by...
870

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関連する実験動画

Updated: May 3, 2026

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
11:36

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing

Published on: July 3, 2016

11.3K

NMR誘導変異を用いたタンパク質の液体-液体相分離を調節するためのプロトコル.

Mayu Enomoto-Kusano1, Kyoko Furuita2,3, Takashi S Kodama2

  • 1Graduate School of Engineering Science, Yokohama National University, Tokiwadai 79-5, Hodogaya-ku, Yokohama 240-8501, Kanagawa, Japan.

Methods and protocols
|February 20, 2026
PubMed
まとめ

研究者は,タンパク質構造と液体液相分離 (LLPS) を結びつける新しい方法を開発しました. このテクニックは,核磁共鳴 (NMR) と突然変異を用いてタンパク質の振る舞いを制御し,細胞区画形成の洞察を提供します.

キーワード:
NMRによる誘導性変異遺伝.VAPBは,VAPBと一致している.液体液体相分離装置核磁気共鳴による核磁気共鳴は,タンパク質コンデンサートは,タンパク質のダイナミクス

さらに関連する動画

A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates
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A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates

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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

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関連する実験動画

Last Updated: May 3, 2026

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
11:36

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing

Published on: July 3, 2016

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A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates
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A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates

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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

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科学分野:

  • バイオケミストリーと分子生物学
  • 細胞生物学 細胞生物学
  • バイオフィジックス 生物物理学

背景:

  • 膜のない細胞区は,液体-液体相分離 (LLPS) によって形成されます.
  • 定量的なLLPSの振る舞いを残留レベル構造データと関連付ける実験方法は限られている.
  • LLPSの分子基盤を理解することは,細胞生物学にとって極めて重要です.

研究 の 目的:

  • 定量的なLLPS測定を,残留レベル構造情報と接続する統合プロトコルを開発する.
  • 構造誘導型ミュータゲネシスによるタンパク質相分離の調節を可能にする.
  • タンパク質LLPS.の系統的,残留レベル制御のための一般化可能な枠組みを提供すること.

主な方法:

  • 核磁共振 (NMR) スペクトロスコーピーの組み合わせた定量的なLLPSアッセイ.
  • タンパク質の配列を修正するために,構造誘導型変異を生成する.
  • VAPB MSPドメインをモデルシステムとして利用した.

主要な成果:

  • 残留物特有の構造的特徴をマクロスコピック LLPS 行動と成功裏に結びつけました.
  • 標的型アミノ酸置換によるタンパク質相分離の抑制と強化が実証されています.
  • LLPS規制を研究するためのモデルとしてVAPB MSPドメインを検証しました.

結論:

  • 開発されたプロトコルは,LLPSの分子決定因子を解剖するための強力なツールを提供します.
  • このフレームワークは,タンパク質相分離の精密な残留レベル操作を可能にします.
  • この発見は,細胞区画形成を理解し,制御するための新しい道を開く.