Related Experiment Video
Updated: Mar 6, 2026
11:39
Identification of Functional Protein Regions Through Chimeric Protein Construction
Published on: January 8, 2019
11.0K
Construction of Protein Switches by Domain Insertion and Directed Evolution
Lucas F Ribeiro1, Tiana D Warren1, Marc Ostermeier2
1Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 N. Charles St, Baltimore, MD, 21218, USA.
Methods in Molecular Biology (Clifton, N.J.)
|March 16, 2017
Summary
Researchers created novel protein switches by randomly fusing protein domains. These switches change between active and inactive states, enabling new biosensor and therapeutic applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Synthetic Biology
Background:
- Protein switches are essential biological components that modulate cellular processes.
- These switches transition between inactive and active states, responding to specific signals.
- Engineering protein switches offers potential for novel biotechnological tools.
Purpose of the Study:
- To develop methods for creating novel protein switches.
- To enable the identification of functional protein switches through selection and screening.
Main Methods:
- Randomly fusing two protein domains to generate diverse libraries.
- Utilizing domain insertion strategies for protein engineering.
- Employing selection and screening techniques to identify functional switches.
Main Results:
- Successful generation of domain insertion libraries.
- Demonstrated feasibility of identifying protein switches from these libraries.
- Established a foundation for creating custom-designed protein switches.
Conclusions:
- Random domain fusion is an effective strategy for protein switch discovery.
- Domain insertion libraries provide a powerful platform for protein engineering.
- This work facilitates the development of novel biosensors and therapeutics.
Related Concept Videos
02:26Conservation of Protein Domains Over Different Proteins
14.9K
Protein domains are small structurally independent units that are part of a single amino acid chain. Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
14.9K
01:49Conserved Binding Sites
5.2K
Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
5.2K
01:57Protein Complexes with Interchangeable Parts
3.0K
Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
3.0K
02:53Conservative Site-specific Recombination and Phase Variation
7.1K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
7.1K
02:26Conservation of Protein Domains
4.3K
4.3K
01:25Protein Folding
12.0K
Proteins are chains of amino acids linked together by peptide bonds. Upon synthesis, a protein folds into a three-dimensional conformation, critical to its biological function. Interactions between its constituent amino acids guide protein folding, and hence the protein structure is primarily dependent on its amino acid sequence.
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
12.0K