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¹H NMR of Labile Protons: Deuterium (²H) Substitution00:48

¹H NMR of Labile Protons: Deuterium (²H) Substitution

This lesson illustrates the role of deuterium substitution in simplifying the NMR spectrum of compounds comprising labile protons. One method employed is the use of deuterium. Amongst the three isotopes of hydrogen, deuterium (2H) has a nucleus composed of one proton and one neutron. When the D2O solvent is added to a pure dry ethanol solution, its labile proton is substituted with deuterium.
¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)01:20

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

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...
Two-Dimensional (2D) NMR: Overview01:12

Two-Dimensional (2D) NMR: Overview

The 1D NMR spectrum of large and complex molecules like natural products has complicated splitting patterns and overlapping signals, which can be easily interpreted using 2-dimensional (2D) NMR. Unlike 1D NMR, 2D NMR has two frequency axes that provide the coupling information between the nucleus A and nucleus B in a molecule. The process from which 2D spectra are obtained has four steps.
The first step is the preparation period, during which nucleus A is excited with a radiofrequency pulse.
¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR01:15

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

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.
Applications Of NMR In Biology01:25

Applications Of NMR In Biology

Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics  for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.
The...
Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

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...

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関連する実験動画

Updated: Jul 16, 2026

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy
14:55

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy

Published on: September 17, 2017

大分子および超分子構造のための溶液NMR技術

Roland Riek1, Jocelyne Fiaux, Eric B Bertelsen

  • 1Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule Zürich, CH-8093 Zürich, Switzerland.

Journal of the American Chemical Society
|October 10, 2002
PubMed
まとめ

この研究は,2D NMRを用いた大規模なバイオ分子分析のために,極化移転技術と横断リラックス最適化スペクトルスコピー (TROSY) を組み合わせた研究である. 最適化された実験パラメータにより,最大800 kDa のマクロモレキュルの高解像度スペクトルが可能です.

科学分野:

  • 生物物理化学 生物物理化学
  • 構造生物学 構造生物学とは
  • 核磁共振 (NMR) スペクトロスコピー

背景:

  • 溶液NMRを用いた大型ホモオリゴメリックマクロ分子 (110~800 kDa) の分析は,信号拡大による課題を提示する.
  • 標準的なNMR技術は,このような大きな生物学的構造に高解像度スペクトルを提供するのにしばしば苦労します.

研究 の 目的:

  • 大型 (15) N,(2) Hラベル付ホモオリゴメリックマクロモレキュルの高品質の相関スペクトルを得るために2D NMR方法を開発し,最適化します.
  • 大型のバイオ分子構造に対するTROSYとCRIPT/CRINEPTの組み合わせの性能を調査する.

主な方法:

  • クロス・コレレート・リラクゼーション・インダクテッド・ポラライゼーション・トランスファー (CRIPT) またはクロス・コレレート・リラクゼーション・エンハンスド・ポラライゼーション・トランスファー (CRINEPT) と組み合わせた横断リラクゼーション・最適化スペクトロスコーピー (TROSY).
  • 取得した2D溶液NMR相関スペクトルは,110から800kDaまでの (15) N,(2) Hラベル付きホモオリゴメリックマクロモレキュルのものです.
  • 系統的に最適化された極化移転時間,リラックス延期,水処理ルーチン.

主要な成果:

  • 管理可能なライン幅 (例えば,800 kDa で 15N の ~75 Hz) を有する TROSY ベースのスペクトルを達成しました.

さらに関連する動画

Disentangling Glycan-Protein Interactions: Nuclear Magnetic Resonance (NMR) to the Rescue
07:40

Disentangling Glycan-Protein Interactions: Nuclear Magnetic Resonance (NMR) to the Rescue

Published on: May 17, 2024

Using Solution NMR to Characterize Biomolecular Condensates Under Biphasic Conditions
09:01

Using Solution NMR to Characterize Biomolecular Condensates Under Biphasic Conditions

Published on: April 17, 2026

関連する実験動画

Last Updated: Jul 16, 2026

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy
14:55

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy

Published on: September 17, 2017

Disentangling Glycan-Protein Interactions: Nuclear Magnetic Resonance (NMR) to the Rescue
07:40

Disentangling Glycan-Protein Interactions: Nuclear Magnetic Resonance (NMR) to the Rescue

Published on: May 17, 2024

Using Solution NMR to Characterize Biomolecular Condensates Under Biphasic Conditions
09:01

Using Solution NMR to Characterize Biomolecular Condensates Under Biphasic Conditions

Published on: April 17, 2026

  • 分子サイズに逆比例する最適な偏振移転時間 (例えば,800 kDa に対して 1.4 ms) を決定する.
  • H2O中の素早い陽子の縦断的なリラックスにより,短いリサイクルタイム (<1s) が確立されています.

    • 結論:

    • 組み合わせたCRIPT/CRINEPT-TROSYアプローチは,大型ホモオリゴメリックマクロモレキュルの高解像度2D NMRスペクトルを効果的に得ることができます.
    • 実験パラメータの最適化,特に転送時間と水圧抑制は,大規模な構造物の成功的なNMR分析に不可欠です.
    • 開発された方法は,溶液中の大きな生物分子組成物のNMR研究を大幅に前進させています.