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

You might also read

Related Articles

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

Sort by
Same author

Ecto-NOX Disulfide-Thiol Exchanger 2 (ENOX2/tNOX) Is a Potential Prognostic Marker in Primary Malignant Melanoma and May Serve as a Therapeutic Target.

International journal of molecular sciences·2024
Same author

The Proteolytic Activity of Neutrophil-Derived Serine Proteases Bound to the Cell Surface Arming Lung Epithelial Cells for Viral Defense.

Molecules (Basel, Switzerland)·2024
Same author

Diagnostic performance of rapid antigen testing for SARS-CoV-2: the COVid-19 AntiGen (COVAG) extension study.

Frontiers in medicine·2024
Same author

Evidence for direct interaction between the oncogenic proteins E6 and E7 of high-risk human papillomavirus (HPV).

The Journal of biological chemistry·2023
Same author

From an Hsp90 - binding protein to a peptide drug.

microLife·2023
Same author

Variation of Proteolytic Cleavage Sites towards the N-Terminal End of the S2 Subunit of the Novel SARS-CoV-2 Omicron Sublineage BA.2.12.1.

Molecules (Basel, Switzerland)·2022

Related Experiment Video

Updated: Jun 28, 2026

Monitoring Neutrophil Elastase and Cathepsin G Activity in Human Sputum Samples
09:23

Monitoring Neutrophil Elastase and Cathepsin G Activity in Human Sputum Samples

Published on: May 21, 2021

Quantifying cathepsin S activity in antigen presenting cells using a novel specific substrate.

Nicolas Lützner1, Hubert Kalbacher

  • 1Interfaculty Institute of Biochemistry, Medical and Natural Sciences Research Centre, University of Tuebingen, Tuebingen, Germany.

The Journal of Biological Chemistry
|October 30, 2008
PubMed
Summary

Researchers developed specific peptide substrates to measure Cathepsin S (CatS) activity in antigen-presenting cells (APCs). This allows for precise quantification of CatS in different APC types and cellular compartments without inhibitors.

More Related Videos

A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl
05:20

A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl

Published on: November 28, 2014

Quantification of Proliferating Human Antigen-specific CD4+ T Cells using Carboxyfluorescein Succinimidyl Ester
07:00

Quantification of Proliferating Human Antigen-specific CD4+ T Cells using Carboxyfluorescein Succinimidyl Ester

Published on: June 4, 2019

Related Experiment Videos

Last Updated: Jun 28, 2026

Monitoring Neutrophil Elastase and Cathepsin G Activity in Human Sputum Samples
09:23

Monitoring Neutrophil Elastase and Cathepsin G Activity in Human Sputum Samples

Published on: May 21, 2021

A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl
05:20

A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl

Published on: November 28, 2014

Quantification of Proliferating Human Antigen-specific CD4+ T Cells using Carboxyfluorescein Succinimidyl Ester
07:00

Quantification of Proliferating Human Antigen-specific CD4+ T Cells using Carboxyfluorescein Succinimidyl Ester

Published on: June 4, 2019

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • Cathepsin S (CatS) is a lysosomal cysteine protease in the papain superfamily.
  • The broad substrate specificity of this enzyme family has hindered the development of specific substrates for CatS.
  • Accurate measurement of CatS activity is crucial for understanding its role in antigen-presenting cells (APCs).

Purpose of the Study:

  • To design and synthesize specific peptide substrates for Cathepsin S (CatS).
  • To enable the quantification of CatS activity in antigen-presenting cells (APCs) without the need for protease inhibitors.
  • To investigate variations in CatS activity across different APC types and subcellular compartments.

Main Methods:

  • Systematic modification of peptide sequences based on GRWHTVGLRWE-Lys(Dnp)-DArg-NH2 to identify CatS cleavage specificity.
  • Synthesis of modified substrates, including Mca-GRWPPMGLPWE-Lys(Dnp)-DArg-NH2 and Mca-GRWHPMGAPWE-Lys(Dnp)-DArg-NH2.
  • Testing substrate specificity against purified cysteine proteases (CatB, CatL) and in endosomal fractions of APCs using CatS inhibitors.

Main Results:

  • Cathepsin S specificity was primarily determined by the P-2, P-1', and P-3' substrate positions.
  • Two novel substrates specifically reacted with CatS and not with CatB or CatL.
  • These substrates allowed for specific quantification of CatS activity in APCs (B cells, macrophages, dendritic cells) without inhibitors.
  • Significant differences in CatS activity were observed between APC types and between lysosomal and endosomal compartments.

Conclusions:

  • Novel, specific peptide substrates for Cathepsin S have been successfully developed.
  • These substrates enable precise and inhibitor-free measurement of CatS activity in various APCs and cellular compartments.
  • The findings highlight distinct CatS activity profiles in different APCs and subcellular locations, suggesting specialized roles.