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Related Concept Videos

Allosteric Regulation01:08

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...
Allosteric Regulation01:08

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...
Ligand Binding and Linkage00:49

Ligand Binding and Linkage

Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked.  In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence the...
Ligand Binding and Linkage00:49

Ligand Binding and Linkage

Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked.  In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence the...
Cooperative Allosteric Transitions01:58

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...
Cooperative Allosteric Transitions01:58

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...

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Related Experiment Video

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Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation
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Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation

Published on: October 4, 2024

Allosteric control of ligand-binding affinity using engineered conformation-specific effector proteins.

Shahir S Rizk1, Marcin Paduch, John H Heithaus

  • 1Department of Biochemistry and Molecular Biology and the Institute for Biophysical Dynamics, University of Chicago, Chicago, Illinois, USA.

Nature Structural & Molecular Biology
|March 8, 2011
PubMed
Summary

Researchers engineered synthetic antigen binders (sABs) to control protein function by targeting specific protein shapes. These sABs enhance maltose binding, offering a new way to modulate biological processes.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Engineering

Background:

  • Maltose-binding protein (MBP) undergoes conformational changes upon ligand binding.
  • Allosteric regulation is crucial for modulating protein function.
  • Engineering proteins to target specific conformations remains a challenge.

Purpose of the Study:

  • To develop a phage display methodology for engineering synthetic antigen binders (sABs).
  • To create sABs that recognize specific conformations of maltose-binding protein (MBP).
  • To investigate the allosteric effects of these sABs on MBP's ligand-binding affinity.

Main Methods:

  • Phage display for library construction and screening.
  • Crystallography for structural analysis of sAB-MBP complexes.
  • In vitro and in vivo assays to assess maltose binding and bacterial growth.

Main Results:

  • Successfully engineered sABs targeting apo and ligand-bound MBP conformations.
  • sABs recognizing the maltose-bound MBP acted as positive allosteric effectors, increasing maltose affinity.
  • Crystal structure revealed the mechanism of allosteric modulation.
  • sABs rescued binding-deficient MBP mutants and enhanced maltose binding in vivo, conferring a growth advantage.

Conclusions:

  • Structure-specific sABs can be engineered to dynamically control ligand-binding affinities.
  • Targeting protein conformations offers a novel strategy for allosteric modulation.
  • This approach has potential applications in biotechnology and therapeutic development.