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Related Concept Videos

Ligand Binding Sites02:40

Ligand Binding Sites

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Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
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A simple method for developing lysine targeted covalent protein reagents.

Ronen Gabizon1, Barr Tivon1, Rambabu N Reddi1

  • 1Department of Chemical and Structural Biology, The Weizmann Institute of Science, Rehovot, 7610001, Israel.

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|December 1, 2023
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Summary
This summary is machine-generated.

Researchers developed versatile thio-methacrylate esters to convert peptides and proteins into potent covalent binders. This method enables targeting of shallow protein surfaces, offering a simple route for irreversible protein labeling and enhanced stability.

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Area of Science:

  • Chemical Biology
  • Biochemistry
  • Drug Discovery

Background:

  • Small molecules struggle to target shallow protein surfaces.
  • Peptide-based covalent probes offer potential but require versatile modification strategies.
  • Targeting broad residue ranges on proteins is challenging.

Purpose of the Study:

  • To develop a versatile method for converting peptides and proteins into covalent binders.
  • To enable targeting of shallow protein surfaces and a broad range of residues.
  • To create potent and stable protein-based covalent binders.

Main Methods:

  • Installation of thio-methacrylate ester-electrophiles on unprotected peptides and proteins via cysteine side chains.
  • Selective reaction of these electrophiles with cysteine and lysine side chains on target proteins.
  • Application of methacrylate-modified peptides and proteins to irreversibly label target proteins.

Main Results:

  • Methacrylate phosphopeptides irreversibly labeled 14-3-3σ via lysine or cysteine residues.
  • Methacrylate peptides targeting lysine showed pan-isoform binding of 14-3-3 proteins.
  • A methacrylate-modified immunity protein irreversibly bound its target DNAse, increasing complex stability.

Conclusions:

  • The thio-methacrylate ester approach provides a simple and versatile method for creating peptide and protein-based covalent binders.
  • This strategy allows for efficient and selective covalent modification of target proteins.
  • The developed covalent binders exhibit potent activity and enhanced stability, with potential applications in drug discovery.