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 Experiment Videos

Salts and glycine increase reversibility and decrease aggregation during thermal unfolding of ribonuclease-A.

Yoshiko Kita1, Tsutomu Arakawa

  • 1Alliance Protein Laboratories, Thousand Oaks, CA 91360, USA.

Bioscience, Biotechnology, and Biochemistry
|May 31, 2002
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Editorial for Special Issue "Recombinant Proteins for Molecular Biology Research: Technologies and Applications".

Current issues in molecular biology·2026
Same author

Parameter optimization for circular dichroism spectroscopy of proteins: A practical approach for rapid acquisition of high-quality spectra.

Analytical biochemistry·2026
Same author

A robust and sensitive method for detecting subtle structural differences in bovine serum albumin.

BioTechniques·2026
Same author

Effects of Arginine on Hierarchical Protein Aggregation.

The protein journal·2026
Same author

Differences and Similarities in Protein and Nucleic Acid Structures and Their Biological Interactions.

Current issues in molecular biology·2026
Same author

Non-ionic Detergent-Assisted Refolding of Protein from Protein-SDS Complex.

The protein journal·2025

Protein unfolding studies of Ribonuclease-A (RNase-A) are complicated by aggregation. Salts and glycine enhance RNase-A reversibility and reduce aggregation during thermal unfolding.

Area of Science:

  • Biochemistry
  • Protein dynamics
  • Biophysical chemistry

Background:

  • Ribonuclease-A (RNase-A) is a widely used model for protein folding and unfolding studies.
  • Protein unfolding reversibility is crucial for understanding protein stability and function.
  • Aggregation can complicate the interpretation of unfolding experiments.

Purpose of the Study:

  • To investigate the effect of aggregation on the thermal unfolding reversibility of RNase-A at neutral pH.
  • To evaluate the impact of additives like salts and glycine on RNase-A aggregation and unfolding reversibility.

Main Methods:

  • Circular dichroism thermal scans to assess protein melting temperature and reversibility.
  • Native-polyacrylamide gel electrophoresis (Native-PAGE) to detect soluble oligomer formation.

Related Experiment Videos

  • Utilized additives such as ammonium sulfate, glycine, and sodium chloride (NaCl).

    • Main Results:

    • RNase-A unfolding at neutral pH showed limited reversibility (approximately 63%) due to aggregation.
    • Heating RNase-A at 75°C resulted in the formation of soluble oligomers.
    • 0.4 M ammonium sulfate increased the melting temperature by ~3°C and improved reversibility.
    • 0.4 M glycine and NaCl significantly enhanced reversibility and reduced aggregation without altering the melting temperature.

    Conclusions:

    • Aggregation significantly contributes to the partial irreversibility of RNase-A thermal unfolding.
    • Salts and glycine are effective in mitigating aggregation and promoting reversibility during RNase-A thermal unfolding.
    • These findings highlight the importance of considering aggregation in protein unfolding studies and suggest strategies for enhancing protein refolding.