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

Conservation of Protein Domains Over Different Proteins02:26

Conservation of Protein Domains Over Different Proteins

10.9K
Protein domains are small structurally independent units that are part of a single amino acid chain.  Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
10.9K
Molecular Chaperones and Protein Folding03:00

Molecular Chaperones and Protein Folding

18.0K
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...
18.0K
Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

2.5K
Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
2.5K
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

5.8K
Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
5.8K
Conserved Binding Sites01:49

Conserved Binding Sites

4.2K
Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
4.2K

You might also read

Related Articles

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

Sort by
Same author

SIRT2-mediated deacetylation activates USP22 catalytic function for PD-L1 protein stabilization and tumor immune escape.

The Journal of clinical investigation·2026
Same author

NRP1 regulates basal nociception: characterization of a peptidergic-specific NRP1 knockout mouse.

bioRxiv : the preprint server for biology·2026
Same author

Beyond linear transcripts: circular RNAs as sentinels of opioid-induced maladaptation in human nociceptors.

Pain·2026
Same author

A novel S1P analogue/MLCK inhibitory peptide-encargoed nanocarrier to attenuate lung vascular leak.

American journal of respiratory cell and molecular biology·2026
Same author

Natural Products Inspired Scaffold Diversification Leads to Unnatural Molecular Warhead and Covalent Strategy to Modulating Protein Function through Electrophilic Bromine Transfer.

Advanced science (Weinheim, Baden-Wurttemberg, Germany)·2026
Same author

GRP78 Selective Inhibitors From a Direct-to-Biology Strategy.

Angewandte Chemie (International ed. in English)·2026
Same journal

Aromatic Cage-Directed Azide-Methyllysine Photochemistry for Profiling Nonhistone Interacting Partners of the MeCP2 Methyl-CpG-Binding Domain.

Biochemistry·2026
Same journal

Differential Hydroxypyruvate Processing by <i>E. coli</i> and <i>P. aeruginosa</i> DXP Synthases Reveals Preferential Xylulose 5-Phosphate Formation by the <i>P. aeruginosa</i> Enzyme.

Biochemistry·2026
Same journal

Structural and Functional Characterization of Heterologous Nitrogenase Complexes.

Biochemistry·2026
Same journal

Discovery of Bacterial Unspecific Peroxygenases.

Biochemistry·2026
Same journal

Lactate Biology: Subcellular Routing and Chemical Form Define Function.

Biochemistry·2026
Same journal

Nature's Anaerobic Toolkit: Glycyl Radical Enzymes and Their Expanding Functional and Mechanistic Diversity.

Biochemistry·2026
See all related articles

Related Experiment Video

Updated: Jul 5, 2025

In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells
00:08

In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells

Published on: September 2, 2019

7.1K

Human Hsp70 Substrate-Binding Domains Recognize Distinct Client Proteins.

Andrew J Ambrose1, Christopher J Zerio1, Jared Sivinski1

  • 1Department of Pharmacology and Toxicology, University of Arizona, Tucson, Arizona 85721, United States.

Biochemistry
|January 20, 2024
PubMed
Summary
This summary is machine-generated.

Human heat shock protein 70 (Hsp70) isoforms exhibit distinct substrate specificities, challenging the role of J-domain containing proteins (JDPs) as the sole drivers of diversity. This finding impacts therapeutic strategies for cancer and neurodegenerative diseases.

More Related Videos

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
11:37

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry

Published on: November 29, 2013

18.5K
Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
06:51

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

Published on: July 21, 2021

2.8K

Related Experiment Videos

Last Updated: Jul 5, 2025

In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells
00:08

In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells

Published on: September 2, 2019

7.1K
Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
11:37

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry

Published on: November 29, 2013

18.5K
Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
06:51

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

Published on: July 21, 2021

2.8K

Area of Science:

  • Molecular Biology
  • Protein Biochemistry
  • Cellular Stress Response

Background:

  • Human cells possess 13 Hsp70 proteins, with substrate specificity often attributed to J-domain containing protein (JDP) cofactors.
  • Previous understanding suggested JDPs primarily dictate Hsp70 substrate binding diversity.

Purpose of the Study:

  • To investigate substrate specificity among the 8 canonical human Hsp70 isoforms.
  • To determine if Hsp70s possess inherent substrate recognition capabilities independent of JDPs.

Main Methods:

  • Peptide arrays were employed to characterize binding affinities between Hsp70 isoforms and peptide substrates.
  • Machine learning algorithms were developed to predict Hsp70-binding sequences based on peptide array data.
  • In vivo complementation assays using *Escherichia coli* DnaK knockout strains and chimeric Hsp70s were performed.

Main Results:

  • Drastically different binding affinities were observed among Hsp70 isoforms for specific peptide substrates.
  • Machine learning revealed novel Hsp70 recognition patterns not predictable by simple sequence alignment.
  • Human Hsp70 isoforms could not complement heat-shocked *E. coli* DnaK knockout cells, but chimeric Hsp70s with DnaK's substrate-binding domain could.
  • Differences in complementation were not linked to DnaJ binding.

Conclusions:

  • Human Hsp70 isoforms possess intrinsic substrate specificity, independent of JDPs.
  • The substrate-binding domain of Hsp70 plays a crucial role in determining in vivo function.
  • Understanding Hsp70 isoform specificity is critical for developing targeted therapies for cancer and neurodegeneration.