Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Molecular Chaperones and Protein Folding

Molecular Chaperones and Protein Folding

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...

Complementation Tests

Complementation Tests

A complementation test is a simple cross to identify whether the two mutations are located on the same gene or different genes. It was first performed by Edward Lewis in the 1940s while working on fruit flies. He developed the test to identify the location and arrangement of different mutations on chromosomes.
Organisms heterozygous for different mutations are crossed pairwise in all combinations. If present on different genes, the mutations can complement each other by providing the missing...

Bacterial Protein Maturation

Bacterial Protein Maturation

Bacterial protein maturation is a tightly regulated process that ensures newly synthesized polypeptides achieve correct functional conformations. This maturation involves a series of modifications, folding events, and quality control steps, often assisted by specialized chaperone proteins.N-Terminal ModificationsThe maturation of bacterial polypeptides begins cotranslationally as the polypeptide exits the ribosome. The first amino acid, N-formylmethionine (fMet), is typically modified at the...

You might also read

Related Articles

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

Sort by

Same author

Snippets from the 28th Congress of the South African Society for Biochemistry and Molecular Biology.

Advances in experimental medicine and biology·2026

Same author

Malaria Public Health Status: Global Context and Update on Gulf Cooperation Council Countries.

Advances in experimental medicine and biology·2026

Same author

Native MS and ligand observed NMR uncovers subtle SLiM binding variations that mediate HSP90-Hop PPI modulation.

RSC chemical biology·2026

Same author

<i>Plasmodium knowlesi</i> Heat Shock Protein 90s: <i>In Silico</i> Analysis Reveals Unique Druggable Structural Features.

International journal of molecular sciences·2025

Same author

J-domain proteins: from molecular mechanisms to diseases.

Cell stress & chaperones·2025

Same author

Genetic Variation in Drug Targets: Are We Ready for the Era of Precision Medicinal Chemistry?

ACS medicinal chemistry letters·2025

Same journal

Nanotechnology-Stem Cell Strategies in 3D Glioblastoma Organoid: Targeting Glioma Stem Cells Within a Complex Tumor Microenvironment.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Mapping the 3D Chromosome Organization of a Biosynthetic Gene Cluster by Capture Hi-C (CHi-C).

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Mapping the 3D Chromosome Organization of Streptomyces by Hi-C.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

CUT&Tag Epigenomic Profiling of Biosynthetic Gene Clusters in Arabidopsis thaliana.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Rhizobium rhizogenes-Mediated Hairy Root Transformation Protocol for Lotus japonicus and Other Legumes.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Characterization of Bioactive Saponins from Sea Cucumbers.

Methods in molecular biology (Clifton, N.J.)·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jul 20, 2025

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Published on: July 21, 2021

Complementation Assays for Co-chaperone Function.

Adrienne L Edkins¹, Gregory L Blatch^2,3,4

¹Biomedical Biotechnology Research Unit (BioBRU), Department of Biochemistry and Microbiology, Rhodes University, Makhanda, South Africa. a.edkins@ru.ac.za.

Methods in Molecular Biology (Clifton, N.J.)

|August 4, 2023

Summary

This summary is machine-generated.

Researchers used mutant microorganisms to test the function of J domain proteins (JDPs), also known as Hsp40s. This method successfully characterized JDPs from bacteria and parasites, advancing co-chaperone research.

Keywords:

DnaJ Heat shock proteins Hsp40 J domain proteins Molecular chaperones Protein folding

More Related Videos

Author Spotlight: Exploring Heat Shock Proteins in Malaria and Tuberculosis Infections

Author Spotlight: Exploring Heat Shock Proteins in Malaria and Tuberculosis Infections

Published on: March 8, 2024

Using Caenorhabditis elegans to Screen for Tissue-Specific Chaperone Interactions

Using Caenorhabditis elegans to Screen for Tissue-Specific Chaperone Interactions

Published on: June 7, 2020

Related Experiment Videos

Last Updated: Jul 20, 2025

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Published on: July 21, 2021

Author Spotlight: Exploring Heat Shock Proteins in Malaria and Tuberculosis Infections

Author Spotlight: Exploring Heat Shock Proteins in Malaria and Tuberculosis Infections

Published on: March 8, 2024

Using Caenorhabditis elegans to Screen for Tissue-Specific Chaperone Interactions

Using Caenorhabditis elegans to Screen for Tissue-Specific Chaperone Interactions

Published on: June 7, 2020

Area of Science:

Molecular Biology
Microbiology
Biochemistry

Background:

J domain proteins (JDPs), formerly Hsp40s, act as co-chaperones essential for protein folding and cellular stress response.
Mutant microorganisms lacking functional JDPs exhibit stress-sensitive phenotypes, providing a basis for functional complementation assays.
Complementation assays allow for the in vivo assessment of novel JDPs by their ability to rescue these stress phenotypes.

Purpose of the Study:

To outline and demonstrate the utility of thermosensitive mutant microorganisms in functional complementation assays for JDPs.
To functionally characterize a JDP from Agrobacterium tumefaciens (AgtDnaJ) and another from Trypanosoma cruzi (TcJ2) using these assays.

Main Methods:

Utilized thermosensitive mutant strains of Escherichia coli (OD259) and Saccharomyces cerevisiae (JJ160) lacking essential JDPs.
Exogenously expressed novel JDPs (AgtDnaJ and TcJ2) in these mutant strains.
Assessed the ability of the expressed JDPs to complement the thermosensitive phenotype, indicating functional co-chaperone activity.

Main Results:

Demonstrated successful functional complementation of thermosensitive phenotypes in both bacterial and yeast mutant strains.
Confirmed the in vivo co-chaperone activity of Agrobacterium tumefaciens JDP (AgtDnaJ) and Trypanosoma cruzi JDP (TcJ2).
Validated the use of these specific thermosensitive microorganisms as effective tools for characterizing JDP function.

Conclusions:

Thermosensitive mutant microorganisms provide a robust platform for the functional characterization of JDPs from diverse organisms.
The study successfully validated the co-chaperone function of AgtDnaJ and TcJ2, contributing to our understanding of JDPs in different biological contexts.
This methodology facilitates the discovery and functional analysis of novel JDPs, crucial for understanding cellular protein homeostasis.