Related Experiment Video
Updated: May 22, 2026
08:06
The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions
Published on: February 1, 2018
Engineered regulation of lysozyme by the SH3-CB1 binding interaction
1Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario, Canada M5S 3G9.
Protein Engineering, Design & Selection : PEDS
|April 26, 2012
Summary
Researchers engineered a synthetic protein switch controlling lysozyme activity using structural design. This protein engineering approach enables precise control over enzyme function via peptide-domain interactions.
Area of Science:
- Protein engineering
- Structural biology
- Biochemistry
Background:
- Protein structural information is key for designing novel therapeutics and diagnostics.
- Controlling protein activity is crucial for biological regulation and synthetic biology.
Purpose of the Study:
- To design and validate a synthetic protein switch using protein structural information.
- To demonstrate control over lysozyme activity via steric hindrance mediated by peptide-domain binding.
Main Methods:
- Computational modeling of fusion proteins (lysozyme-CB1) to predict steric hindrance.
- In vitro synthesis and testing of the designed lysozyme-CB1 fusion protein.
- Assaying lysozyme activity in the presence and absence of the SH3 domain and competitor peptides.
Main Results:
- A lysozyme-CB1 fusion protein was designed and created based on structural modeling.
- The fusion protein exhibited normal lysozyme activity without SH3.
- Lysozyme activity was inhibited by SH3 binding and restored by excess CB1 peptides, demonstrating switch functionality.
Conclusions:
- A structure-based strategy successfully engineered a synthetic protein switch.
- Peptide-domain binding interfaces can be utilized to create synthetic protein regulation.
- This approach holds potential for developing new therapeutic and diagnostic protein products.
More Related Videos
Related Concept Videos
02:58Regulated Protein Degradation
It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
01:28Gene Regulation in Microbial Communities: Quorum Sensing
Quorum sensing is a mechanism of bacterial communication that enables coordinated gene expression in response to changes in population density. This facilitates collective behaviors that enhance survival, resource acquisition, and ecological adaptation. This process relies on small signaling molecules called autoinducers that accumulate as bacterial populations grow. When a critical threshold concentration of autoinducers is reached, bacterial cells collectively modify gene expression,...
01:08Allosteric 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...