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

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution00:52

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution

1.4K
At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the...
1.4K
¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR01:15

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

1.8K
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.
1.8K
Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

1.6K
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...
1.6K
UV–Vis Spectroscopy of Conjugated Systems01:32

UV–Vis Spectroscopy of Conjugated Systems

9.1K
Organic compounds with conjugated double bonds show strong absorption features in the UV–visible region of the electromagnetic spectrum attributed to π → π* electronic excitations. Generally, a UV–vis absorption spectrum is recorded as a plot of absorbance vs wavelength. The wavelength of maximum absorbance, which manifests as a peak in the absorption spectrum, is denoted as λmax.
One of the factors influencing λmax is the extent of conjugation in...
9.1K
¹H NMR: Complex Splitting01:13

¹H NMR: Complex Splitting

2.1K
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...
2.1K
¹H NMR of Labile Protons: Deuterium (²H) Substitution00:48

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

1.5K
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.
1.5K

You might also read

Related Articles

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

Sort by
Same author

A High-Throughput Platform for Measuring and Predicting Vitrification Behavior in Multicomponent Aqueous Solutions.

ACS applied materials & interfaces·2026
Same author

Spatial Organization of Lipids Drives GPCR Conformational Equilibria.

bioRxiv : the preprint server for biology·2026
Same author

Chain Length as a Molecular Determinant of Hydrogen-Bond Dynamics in Biocondensates.

The journal of physical chemistry letters·2026
Same author

An Integrative Biophysical and Computational Workflow Uncovers New Allosteric Sites and Modulators of the Human A<sub>2A</sub> Adenosine Receptor.

ACS chemical biology·2026
Same author

Low-cost calculation and analysis of 2D IR spectra of model diiron trinitrosyl complexes in the NO stretch region with vibrational perturbation theory.

Physical chemistry chemical physics : PCCP·2026
Same author

Membrane Composition Reshapes the Folding Landscape of a pH-Responsive Peptide.

The journal of physical chemistry letters·2025

Related Experiment Video

Updated: Mar 28, 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

16.2K

Solvent Reorganization in Stabilized Protein-Polymer Conjugates Visualized by Two-Dimensional Infrared and Nuclear

Raiza Maia1, Xiaobing Chen1, Emma Mulry2

  • 1Department of Chemistry, University of Texas at Austin, Austin, Texas 78712, United States.

JACS Au
|March 27, 2026
PubMed
Summary

PEGylation enhances biologic stability by stabilizing the protein's solvation shell, not by dehydration. Optimizing polymer length is key for predictable biologic thermal stability.

Keywords:
2D IRGalectin-3MD simulationsNMR spectroscopyPEGylationconformational changesprotein solvation

More Related Videos

Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments
09:25

Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments

Published on: November 1, 2024

3.0K
Author Spotlight: Unveiling the Structural and Dynamic Aspects of Glycan Molecular Recognition
07:40

Author Spotlight: Unveiling the Structural and Dynamic Aspects of Glycan Molecular Recognition

Published on: May 17, 2024

2.1K

Related Experiment Videos

Last Updated: Mar 28, 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

16.2K
Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments
09:25

Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments

Published on: November 1, 2024

3.0K
Author Spotlight: Unveiling the Structural and Dynamic Aspects of Glycan Molecular Recognition
07:40

Author Spotlight: Unveiling the Structural and Dynamic Aspects of Glycan Molecular Recognition

Published on: May 17, 2024

2.1K

Area of Science:

  • Biochemistry
  • Biophysics
  • Materials Science

Background:

  • PEGylation is crucial for biologic function, protecting them from degradation.
  • The exact mechanisms by which PEGylation enhances protein stability, particularly through solvation effects, remain incompletely understood.
  • Understanding these mechanisms is vital for designing more effective PEGylated biologics.

Purpose of the Study:

  • To investigate the role of solvent dynamics in PEGylation-mediated protein stabilization.
  • To elucidate the molecular mechanisms underlying the interaction between PEGylated proteins and their solvent environment.
  • To correlate PEG polymer length with protein dynamics, solvent dynamics, and thermal stability.

Main Methods:

  • Two-dimensional infrared (2D IR) spectroscopy to probe subpicosecond protein and solvent dynamics.
  • Multidimensional nuclear magnetic resonance (NMR) spectroscopy to investigate noncovalent interactions and structural changes.
  • Molecular dynamics (MD) simulations to complement experimental findings and explore solvent accessibility.

Main Results:

  • PEGylation introduces polymer length-dependent differences in protein and solvent dynamics.
  • Slower solvent dynamics were observed to correlate with increased thermal stability of the PEGylated protein.
  • Evidence suggests PEGylation forms a stabilizing 'shroud' around the protein, influencing solvent and backbone dynamics without reducing solvent accessibility.

Conclusions:

  • PEGylation stabilizes proteins by enhancing the stability of the protein's solvation shell, rather than by simple dehydration.
  • The findings challenge the prevailing dehydration hypothesis for PEGylation-mediated stabilization.
  • These insights provide a mechanistic basis for optimizing polymer length to control the thermal stability of biologics.