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 Experiment Videos

The one-bead two-compound assay for solid phase screening of combinatorial libraries.

Morten Meldal1

  • 1Carlsberg Laboratory, Department of Chemistry, Gamle Carlsberg Vej 10, DK-2500 Valby, Denmark.

Biopolymers
|September 27, 2002
PubMed
Summary
This summary is machine-generated.

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

Toward actionable interventions in human aging (12th ARDD meeting, 2025).

Aging·2026
Same author

Click-Cyclized Cell Penetrating Peptides Containing Hydrophobic Proline Derivatives for Efficient Intracellular Delivery.

Angewandte Chemie (International ed. in English)·2025
Same author

Parthenolide disrupts mitosis by inhibiting ZNF207/BUGZ-promoted kinetochore-microtubule attachment.

The EMBO journal·2025
Same author

Green Resins for All: Sustainable Preparation of PEGA Resin for Peptide and Protein Synthesis and Immobilization.

ACS applied materials & interfaces·2025
Same author

Hydrodynamic Control of Alzheimer Aβ Fibrillation with Glucosaminic Acid Containing Click-Cyclized β-Bodies.

Journal of the American Chemical Society·2023
Same author

Cascade autohydrolysis of Alzheimer's Aβ peptides.

Chemical science·2023
Same journal

Untreated Rosehip Powder/Poly(Lactic Acid)/Poly(3-Hydroxybutyrate-Co-4-Hydroxybutyrate) Electrospun Mats for Wound Dressing Applications.

Biopolymers·2026
Same journal

Synthesis, Characterization, and Antidiabetic Evaluation of Sequence-Modified Liraglutide Analogs in a Drosophila melanogaster Model.

Biopolymers·2026
Same journal

Fabrication of an Antibacterial Alginate/Chitosan Hydrogel Dressing Loaded With CuO Nanoparticles for Wound Dressing Applications.

Biopolymers·2026
Same journal

Effect of Chitosan-Alginate Polyelectrolyte Complex Formation and Multilayer Polymer Configuration on the Characteristics of 3D-Printed Metronidazole-Loaded Periodontal Films.

Biopolymers·2026
Same journal

Phenolic Grafting of Oxidized Cellulose Nanofibers Using Ferulic Acid: Structural and Antioxidant Analysis Toward Bioactive Nanomaterials.

Biopolymers·2026
Same journal

Detection of a Target Nucleic Acid by Ligation-Assisted Fluorescence Enhancement of a Peptide Nucleic Acid (PNA) Twin Probe via Disulfide Binding.

Biopolymers·2026
See all related articles

This study presents a novel method using fluorescent quenched substrate libraries to map protease specificity and identify potent enzyme inhibitors. The approach utilizes solid-phase synthesis and bead sorting for efficient screening and characterization of protease activity and inhibition.

Area of Science:

  • Biochemistry
  • Enzymology
  • Chemical Biology

Background:

  • Protease activity and specificity are crucial in biological processes.
  • Investigating proteases requires efficient tools for activity and inhibition screening.
  • Fluorescent resonance energy transfer (FRET) substrates offer a sensitive detection method.

Purpose of the Study:

  • To develop and validate a robust solid-phase assay for mapping protease substrate specificity.
  • To establish a high-throughput method for identifying potent protease inhibitors.
  • To utilize fluorescent quenched substrate libraries for enzyme discovery and characterization.

Main Methods:

  • Preparation of one-bead one-compound libraries using split and combine synthesis.
  • Utilizing poly(ethylene glycol) (PEG)-based resins as solid supports for substrate hydrolysis.

Related Experiment Videos

  • Employing fluorescence microscopy and bead sorting for active substrate and inhibitor isolation.
  • Edman sequencing and mass spectrometry for substrate and inhibitor structure determination.

    • Main Results:

    • Developed a method to generate a complete map of protease substrate specificity.
    • Identified protease inhibitors with high affinity (nM range) using a competitive binding assay.
    • Demonstrated high accuracy with minimal false positives in inhibitor screening.
    • Validated solid-phase findings through resynthesis and solution kinetics.

    Conclusions:

    • Fluorescent quenched substrate libraries on PEG-based resins are effective for protease research.
    • The developed method enables efficient identification of specific substrates and potent inhibitors.
    • This approach significantly advances the study of protease activity, specificity, and inhibition.