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

Protein Folding01:25

Protein Folding

8.8K
Proteins are chains of amino acids linked together by peptide bonds. Upon synthesis, a protein folds into a three-dimensional conformation, critical to its biological function. Interactions between its constituent amino acids guide protein folding, and hence the protein structure is primarily dependent on its amino acid sequence.
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
8.8K
Protein Folding01:22

Protein Folding

112.3K
Overview
112.3K
Protein Folding01:22

Protein Folding

29.7K
29.7K
Molecular Chaperones and Protein Folding03:00

Molecular Chaperones and Protein Folding

14.7K
The native conformation of a protein is formed by interactions between the side chains of its constituent amino acids. When the amino acids cannot form these interactions, the protein cannot fold by itself and needs chaperones. Notably, chaperones do not relay any additional information required for the folding of polypeptides; the native conformation of a protein is determined solely by its amino acid sequence. Chaperones catalyze protein folding without being a part of the folded protein.
The...
14.7K
Molecular Chaperones and Protein Folding03:00

Molecular Chaperones and Protein Folding

14.2K
14.2K
Protein Folding Quality Check in the RER01:29

Protein Folding Quality Check in the RER

4.4K
ER is the primary site for the maturation and folding of soluble and transmembrane secretory proteins. The calnexin cycle is a specific chaperone system that folds and assesses the confirmation of N-glycosylated proteins before they can exit the ER lumen. The primary players of this quality check pipeline are the lectins, ER-resident chaperones, and a glucosyl transferase enzyme. In case the calnexin system in the lumen fails to salvage a misfolded protein, it is transported to the cytoplasm...
4.4K

You might also read

Related Articles

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

Sort by
Same author

LRP4 is an entry receptor for multiple encephalitic alphaviruses.

Nature communications·2026
Same author

CD164 is an endolysomal host factor for entry of Clade A New World Arenaviruses.

bioRxiv : the preprint server for biology·2026
Same author

Protective human antibodies against Powassan virus.

Journal of virology·2026
Same author

Multiple LDLR family members act as entry receptors for yellow fever virus.

Nature·2025
Same author

LRP8 is an entry receptor for tick-borne encephalitis viruses.

Proceedings of the National Academy of Sciences of the United States of America·2025
Same author

Determinants of human versus mosquito cell entry by the Chikungunya virus envelope proteins.

bioRxiv : the preprint server for biology·2025

Related Experiment Video

Updated: May 2, 2026

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins
09:55

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins

Published on: July 4, 2016

12.6K

Oxidative refolding from inclusion bodies.

Christopher A Nelson1, Chung A Lee, Daved H Fremont

  • 1Center for Structural Genomics of Infectious Diseases, Computational Institute, University of Chicago, Chicago, IL, 60557, USA.

Methods in Molecular Biology (Clifton, N.J.)
|March 5, 2014
PubMed
Summary

This study details a method for growing and purifying bacterial inclusion body proteins, including optional selenomethionine labeling. The protocol ensures high-quality protein recovery through denaturation, refolding, and chromatography.

More Related Videos

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain
14:25

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

Published on: December 12, 2017

17.6K
Purification and Refolding to Amyloid Fibrils of His6-tagged Recombinant Shadoo Protein Expressed as Inclusion Bodies in E. coli
09:43

Purification and Refolding to Amyloid Fibrils of His6-tagged Recombinant Shadoo Protein Expressed as Inclusion Bodies in E. coli

Published on: December 19, 2015

13.7K

Related Experiment Videos

Last Updated: May 2, 2026

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins
09:55

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins

Published on: July 4, 2016

12.6K
Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain
14:25

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

Published on: December 12, 2017

17.6K
Purification and Refolding to Amyloid Fibrils of His6-tagged Recombinant Shadoo Protein Expressed as Inclusion Bodies in E. coli
09:43

Purification and Refolding to Amyloid Fibrils of His6-tagged Recombinant Shadoo Protein Expressed as Inclusion Bodies in E. coli

Published on: December 19, 2015

13.7K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Chemistry

Background:

  • Bacterial inclusion bodies are aggregates of misfolded recombinant proteins.
  • Efficient purification and refolding of these proteins are crucial for structural and functional studies.
  • Current methods can be inefficient or require auxotrophic strains.

Purpose of the Study:

  • To describe a robust protocol for the growth and purification of bacterial inclusion body proteins.
  • To provide an optional method for selenomethionine labeling of proteins in common E. coli strains.
  • To ensure high-quality refolded protein suitable for further analysis.

Main Methods:

  • Bacterial expression and induction for inclusion body formation.
  • Chemical denaturation and disulfide reduction for solubilization.
  • Rapid dilution refolding with arginine and glutathione redox buffer.
  • Stirred cell concentration and size-exclusion chromatography for purification.
  • Optional selenomethionine incorporation via methionine biosynthesis inhibition.

Main Results:

  • Successful solubilization and refolding of inclusion body proteins.
  • High recovery of purified, functional recombinant protein.
  • Demonstrated ability to incorporate selenomethionine in non-auxotrophic E. coli.
  • Assessment of protein quality by SDS-PAGE.

Conclusions:

  • The described protocol offers an efficient and versatile method for obtaining high-quality bacterial inclusion body proteins.
  • The optional selenomethionine labeling expands its utility for structural biology applications, particularly X-ray crystallography.
  • This method is applicable to common E. coli strains, simplifying protein production workflows.