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

NMR Spectroscopy Of Amines01:19

NMR Spectroscopy Of Amines

10.9K
In proton NMR spectroscopy, primary amines and secondary amines showcase their N–H protons as a broad signal in the chemical shift range between δ 0.5 and 5 ppm. The exact position in this range depends on several factors, including sample concentration, hydrogen bonding, and the type of solvent used. Since amine protons undergo fast proton exchange in solution, the protons are labile and therefore do not participate in any splitting with adjacent protons. Thus, the observed peak is...
10.9K
¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)01:20

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

1.6K
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...
1.6K
Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

1.3K
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...
1.3K

You might also read

Related Articles

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

Sort by
Same author

Cosolutes Modulate Polyubiquitin Fibrillation.

ACS omega·2026
Same author

Investigating Enzyme-Substrate Interactions in the Assembly of K48/K63 Heterotypic Ubiquitin Chains.

Biochemistry·2026
Same author

Structure-guided engineering of protein stability through core hydrophobicity.

Protein science : a publication of the Protein Society·2025
Same author

Beyond degradation tags: How FAT10 and ubiquitin shape substrate energy landscapes.

Essays in biochemistry·2025
Same author

Ionic Liquid-Induced Modulation of Ubiquitin Stability: The Dominant Role of Hydrophobic Interactions.

Langmuir : the ACS journal of surfaces and colloids·2025
Same author

Structural analysis of genetic variants of the human tumor suppressor PALB2 coiled-coil domain.

Bioscience reports·2025

Related Experiment Video

Updated: Jan 17, 2026

LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics
08:01

LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics

Published on: April 9, 2017

8.5K

Monitoring Protein Deamidation in Real Time Using NMR.

Rashmi Agrata1, Ranabir Das1

  • 1National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India.

Magnetic Resonance in Chemistry : MRC
|September 18, 2025
PubMed
Summary

Bacterial deamidases CIFEC and CIFBP modify host proteins like ubiquitin. CIFBP shows higher catalytic efficiency than CIFEC, offering insights into bacterial pathogenesis and host manipulation mechanisms.

Keywords:
NMRdeamidationreal‐time bacterial effectorsubiquitin

More Related Videos

Methods to Identify the NMR Resonances of the 13C-Dimethyl N-terminal Amine on Reductively Methylated Proteins
13:59

Methods to Identify the NMR Resonances of the 13C-Dimethyl N-terminal Amine on Reductively Methylated Proteins

Published on: December 12, 2013

6.6K
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

3.7K

Related Experiment Videos

Last Updated: Jan 17, 2026

LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics
08:01

LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics

Published on: April 9, 2017

8.5K
Methods to Identify the NMR Resonances of the 13C-Dimethyl N-terminal Amine on Reductively Methylated Proteins
13:59

Methods to Identify the NMR Resonances of the 13C-Dimethyl N-terminal Amine on Reductively Methylated Proteins

Published on: December 12, 2013

6.6K
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

3.7K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Microbiology

Background:

  • Bacterial deamidases are crucial effectors in pathogenesis.
  • They manipulate host signaling by modifying proteins like ubiquitin (Ub) and NEDD8.
  • Understanding their enzymatic activity is key to deciphering bacterial strategies.

Purpose of the Study:

  • To investigate the enzymatic activity of two bacterial deamidases: CIFEC (Escherichia coli) and CIFBP (Burkholderia pseudomallei).
  • To compare the catalytic efficiency of CIFEC and CIFBP using real-time NMR.
  • To explore the utility of NMR spectroscopy for studying deamidation posttranslational modifications.

Main Methods:

  • Utilized traditional and real-time NMR-based techniques.
  • Employed BEST-HSQC NMR spectroscopy for real-time monitoring of ubiquitin deamidation.
  • Performed kinetic analysis to quantify enzyme activity and substrate binding.

Main Results:

  • Demonstrated significant differences in catalytic efficiency between CIFEC and CIFBP.
  • CIFBP exhibited higher catalytic efficiency compared to CIFEC.
  • CIFEC showed weaker substrate binding and suboptimal efficiency, suggesting a regulatory role.

Conclusions:

  • NMR spectroscopy is a robust tool for quantifying deamidation enzyme activity and studying irreversible PTMs.
  • Differences in catalytic efficiency between CIFEC and CIFBP provide mechanistic insights into bacterial pathogenesis.
  • Localized structural changes in ubiquitin upon deamidation warrant further investigation.