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Applications Of NMR In Biology

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.

Chemical Shift: Internal References and Solvent Effects

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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Related Experiment Video

Updated: May 2, 2026

Mapping Molecular Diffusion in the Plasma Membrane by Multiple-Target Tracing MTT

Mapping Molecular Diffusion in the Plasma Membrane by Multiple-Target Tracing MTT

Published on: May 27, 2012

Cell Permeability and Target Engagement of Middle-Sized Molecules Quantified by In-Cell NMR.

Yota Sukigara¹, Hajime Kamoshida², Yuji Tokunaga^1,2

¹Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo, Tokyo 113-0033, Japan.

Analytical Chemistry

|August 25, 2025

Summary

We developed a floating in-cell NMR method to measure how well middle-sized molecules enter cells and engage intracellular targets. This technique offers valuable insights for developing new drugs targeting protein-protein interactions.

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Published on: March 9, 2021

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Last Updated: May 2, 2026

Mapping Molecular Diffusion in the Plasma Membrane by Multiple-Target Tracing MTT

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Published on: May 27, 2012

Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers

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Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor

Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor

Published on: March 9, 2021

Area of Science:

Biochemistry
Molecular Biology
Pharmacology

Background:

Middle-sized molecules, including macrocycles and cyclic peptides, are promising drug candidates for targeting intracellular protein-protein interactions (PPIs).
Assessing cell permeability and intracellular target engagement for these molecules is difficult, with limited available data.
Existing in-cell nuclear magnetic resonance (NMR) methods face challenges with sensitivity and potential artifacts from cell encapsulation.

Purpose of the Study:

To develop a novel floating in-cell NMR strategy for quantifying cell permeability and intracellular target engagement of middle-sized molecules.
To overcome limitations of conventional in-cell NMR methods, enhancing sensitivity and preserving cell viability.
To provide a tool for generating quantitative pharmacodynamic data for drug development targeting intracellular PPIs.

Main Methods:

Developed a floating in-cell NMR technique for live mammalian cells.
Utilized the method to measure cell permeation rates and intracellular target affinities of FK506.
Assessed the applicability of the method to another middle-sized molecule, rapamycin.

Main Results:

The floating in-cell NMR strategy enhanced measurement sensitivity while maintaining cell survival.
Successfully quantified cell permeation and intracellular target engagement for FK506.
Demonstrated that increased temperature accelerates FK506 cell permeation without altering intracellular affinity, indicating temperature-dependent membrane fluidity effects.
Validated the method's applicability to rapamycin.

Conclusions:

The floating in-cell NMR approach provides a sensitive and reliable method for evaluating middle-sized molecule pharmacodynamics in living cells.
This strategy enables quantitative assessment of cell permeability and intracellular target engagement without chemical modification.
The findings offer valuable insights for the rational design of middle-sized molecules targeting intracellular PPIs.