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Related Concept Videos

Enzyme Inhibition01:30

Enzyme Inhibition

Inhibitors are molecules that reduce enzyme activity by binding to the enzyme. In a normally functioning cell, enzymes are regulated by a variety of inhibitors. Drugs and other toxins can also inhibit enzymes. Some inhibitors bind to the enzyme’s active site, while others inhibit enzymatic activity by binding to other sites on the protein structure.
Enzymes02:34

Enzymes

Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...

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Related Experiment Video

Updated: Jun 21, 2026

The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes
09:47

The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes

Published on: May 25, 2018

Achieving protease substrate-specific inhibition by mAb dual functional selections.

Ki Baek Lee1,2, Sangcheon Lee2, Bharati Reddi1

  • 1Texas Therapeutics Institute, Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

Protein Science : a Publication of the Protein Society
|June 20, 2026
PubMed
Summary

This study developed a novel selection method to discover protease inhibitors with substrate specificity. These inhibitors target pathogenic proteolysis while sparing beneficial functions, crucial for developing safer therapeutics.

Keywords:
levansucrasemAbnegative selectionprotease inhibitorsubstrate‐specific inhibition

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Last Updated: Jun 21, 2026

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Area of Science:

  • Biochemistry
  • Enzymology
  • Molecular Biology

Background:

  • Proteases are enzymes with broad substrate activity, making selective inhibition challenging.
  • Developing safe and effective protease inhibitors requires distinguishing between pathogenic and physiological proteolysis.

Purpose of the Study:

  • To establish a dual functional selection system for discovering protease inhibitory monoclonal antibodies (mAbs) with substrate specificity.
  • To isolate and characterize anti-matrix metalloproteinase (MMP)-14 inhibitory mAbs that exhibit substrate-specific inhibition.

Main Methods:

  • Engineered bacterial periplasmic expression systems using beta-lactamase and levansucrase as selection markers.
  • Developed positive and negative selection strategies for identifying protease inhibitory mAbs.
  • Isolated and characterized anti-MMP-14 mAbs for binding affinity, inhibitory potency, selectivity, and substrate specificity.

Main Results:

  • Successfully established a dual functional selection system for protease inhibitor discovery.
  • Isolated anti-MMP-14 inhibitory mAbs with nanomolar affinities and high stability.
  • Demonstrated substrate-specific (SS) inhibition, blocking syndecan-1 cleavage while allowing macrophage chemoattractant protein 3 cleavage.
  • Mechanistic studies revealed active-site competitive inhibition and substrate specificity based on protease subsites.

Conclusions:

  • The developed dual functional selection is effective for discovering substrate-specific protease inhibitors.
  • The isolated anti-MMP-14 mAbs represent promising therapeutic candidates due to their SS inhibition.
  • This selection strategy has broad applicability for targeting other proteases with pharmaceutical significance.