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

Caspases01:24

Caspases

12.4K
Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
12.4K
Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

6.6K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
6.6K
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
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.0K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
6.0K

You might also read

Related Articles

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

Sort by
Same author

Age-Dependent z Scores and eGFR-Adjusted Reference Ranges for Neurofilament Light: A Practical Approach for Clinical Laboratories.

Clinical chemistry·2026
Same author

Unraveling the active site cover of coproheme decarboxylase from Listeria monocytogenes.

The FEBS journal·2026
Same author

Indirect optical geometry measurement based on optical tweezers in transparent microchannels.

Optics express·2026
Same author

Correction: Intraperitoneal Oil Application Causes Local Inflammation with Depletion of Resident Peritoneal Macrophages.

Molecular cancer research : MCR·2026
Same author

Leaky recombinant expression reveals design constraints of bicistronic synthetic operons in Escherichia coli.

Scientific reports·2026
Same author

Identification of serum biomarkers linking myocardial fibrosis, systolic dysfunction and outcomes in patients with severe aortic stenosis.

Cardiovascular research·2026

Related Experiment Video

Updated: Jun 22, 2025

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
19:23

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

Published on: January 16, 2019

9.2K

Caspase-Based Fusion Protein Technology: Substrate Cleavability Described by Computational Modeling and Simulation.

Jakob Liu1,2, Andreas Fischer1,3, Monika Cserjan-Puschmann1,3

  • 1Austrian Centre of Industrial Biotechnology, Muthgasse 18, 1190 Vienna, Austria.

Journal of Chemical Information and Modeling
|July 1, 2024
PubMed
Summary

We developed a method to predict Caspase-based fusion protein technology (CASPON) cleavage efficiency. This approach uses molecular dynamics to analyze protein structures, improving protein purification strategies.

More Related Videos

Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches
05:56

Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches

Published on: October 13, 2022

1.3K
Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation
08:47

Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation

Published on: March 5, 2018

9.0K

Related Experiment Videos

Last Updated: Jun 22, 2025

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
19:23

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

Published on: January 16, 2019

9.2K
Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches
05:56

Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches

Published on: October 13, 2022

1.3K
Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation
08:47

Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation

Published on: March 5, 2018

9.0K

Area of Science:

  • Biochemistry
  • Structural Biology
  • Protein Engineering

Background:

  • Caspase-based fusion protein technology (CASPON) enables universal cleavage of fusion tags to restore native N-termini.
  • CASPON enzyme optimization has yielded promiscuity against various N-terminal peptides.
  • However, cleavage efficiency can be unexpectedly low for larger proteins.

Purpose of the Study:

  • To develop an efficient computational method for predicting CASPON cleavage efficiency.
  • To rationalize cleavage outcomes based on structural interactions between CASPON and N-terminal peptides.

Main Methods:

  • Utilizing molecular dynamics simulations to generate diverse N-terminal peptide conformations.
  • Analyzing the structural representation of intrinsically disordered N-terminal peptides.
  • Employing a fitting procedure to model interactions with the CASPON enzyme.

Main Results:

  • The number of favorable N-terminal conformations strongly correlates with experimentally observed cleavage efficiency.
  • This agreement supports a conformational selection model for CASPON activity.
  • The method provides a computationally inexpensive way to assess cleavability.

Conclusions:

  • A novel computational approach accurately predicts CASPON cleavage efficiency.
  • Understanding N-terminal peptide conformations is key to optimizing protein purification.
  • This method allows for a priori assessment of CASPON cleavability, streamlining protein engineering workflows.