Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

A Cysteine-Dependent Peptide Cyclase with Broad Substrate Tolerance Enables Chemoenzymatic Synthesis of Macolacin Analogs.

ACS chemical biology·2026
Same author

The NMR Exchange Format (NEF): Specification and Applications.

bioRxiv : the preprint server for biology·2026
Same author

Serum Proteomics of Multiple Menstrual Symptoms in Female Athletes: A Pilot Study.

FASEB bioAdvances·2026
Same author

Proteomics to reveal relationship between menstrual cycle and subjective conditions.

Scientific reports·2026
Same author

A Novel Mouse Model of Sympathoadrenal Catecholamine Deficiency by Ablation of the Tyrosine Hydroxylase Gene With Slc6a4-Cre.

Journal of neurochemistry·2026
Same author

Unique Microswitches Positioned Extracellular to the Orthosteric Binding Site Initiate Activation in the β<sub>1</sub>-Adrenergic Receptor.

Journal of chemical information and modeling·2026

Related Experiment Video

Updated: Jun 12, 2026

Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor
10:25

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

NMR protein structure determination in living E. coli cells using nonlinear sampling.

Teppei Ikeya1, Atsuko Sasaki, Daisuke Sakakibara

  • 1Department of Chemistry, Tokyo Metropolitan University, Hachioji, Tokyo, Japan.

Nature Protocols
|June 12, 2010
PubMed
Summary

Researchers developed a new in-cell nuclear magnetic resonance (NMR) method for determining protein structures directly within living cells. This technique enables rapid, high-resolution structural analysis without protein purification, accelerating biochemical research.

More Related Videos

Continuous Measurement of Biological Noise in Escherichia Coli Using Time-lapse Microscopy
08:25

Continuous Measurement of Biological Noise in Escherichia Coli Using Time-lapse Microscopy

Published on: April 27, 2021

Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization
10:01

Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization

Published on: July 30, 2020

Related Experiment Videos

Last Updated: Jun 12, 2026

Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor
10:25

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

Continuous Measurement of Biological Noise in Escherichia Coli Using Time-lapse Microscopy
08:25

Continuous Measurement of Biological Noise in Escherichia Coli Using Time-lapse Microscopy

Published on: April 27, 2021

Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization
10:01

Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization

Published on: July 30, 2020

Area of Science:

  • Biochemistry and Structural Biology
  • Molecular Biophysics
  • Cellular Biology

Background:

  • Cells are crowded environments where proteins interact dynamically.
  • Understanding protein structures in their native cellular context is crucial for biochemical research.
  • Nuclear Magnetic Resonance (NMR) spectroscopy has advanced for macromolecular studies.

Purpose of the Study:

  • To present a protocol for stable isotope labeling and structure determination of proteins within living cells.
  • To demonstrate the feasibility of high-resolution three-dimensional (3D) structure determination using in-cell NMR.
  • To bypass the need for protein purification, enabling faster structural analysis.

Main Methods:

  • Stable isotope labeling ((13)C, (15)N, (2)H) of overexpressed proteins in Escherichia coli.
  • Rapid acquisition of multidimensional NMR spectra using nonlinear sampling.
  • Structure calculation and refinement performed on data obtained exclusively from living cells.

Main Results:

  • Successful determination of the first 3D protein structure (TTHA1718) in living cells using the developed in-cell NMR protocol.
  • Sample preparation and NMR measurement time reduced to 2-3 days and 2-3 hours per experiment, respectively.
  • High-resolution structural information obtained directly from the cellular environment.

Conclusions:

  • The in-cell NMR protocol provides a powerful method for determining protein structures in their native cellular milieu.
  • This approach significantly accelerates structural biology research by eliminating protein purification steps.
  • The method holds promise for understanding protein function and interactions in a biologically relevant context.