Related Experiment Video
Updated: Jun 20, 2026
08:34
OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
Published on: February 5, 2020
Expressed protein ligation: a resourceful tool to study protein structure and function
Luis Berrade1, Julio A Camarero
1Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, 1985 Zonal Avenue, PSC 616, Los Angeles, CA 90033, USA.
Cellular and Molecular Life Sciences : CMLS
|August 18, 2009
Summary
Expressed protein ligation (EPL) enables the creation of precisely defined semi-synthetic proteins. This powerful technique, combined with biochemical analysis, enhances our understanding of protein structure, function, and stability.
Area of Science:
- Biochemistry
- Chemical Biology
- Structural Biology
Background:
- Expressed protein ligation (EPL) is a chemoselective method for joining unprotected polypeptides.
- It enables the synthesis of homogeneous, well-defined semi-synthetic proteins from both synthetic and recombinant sources.
Purpose of the Study:
- To review the applications of EPL in studying protein structure, function, and stability.
- To highlight the increasing sophistication and utility of EPL in addressing complex biological questions.
Main Methods:
- Site-specific introduction of biophysical probes, unnatural amino acids, and post-translational modifications.
- Utilizing EPL for the production of semi-synthetic protein samples.
- Combining EPL with standard biochemical analysis techniques.
Main Results:
- EPL facilitates the creation of homogeneous protein samples with defined chemical compositions.
- The method allows for precise modification of proteins for advanced studies.
- Over the past decade, EPL applications have become more sophisticated.
Conclusions:
- EPL is a powerful technology for protein research.
- Combined with biochemical analysis, EPL significantly improves the understanding of protein structure and function.
- The review underscores the value of EPL in advancing biological inquiry.
Related Concept Videos
02:40Ligand Binding Sites
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...
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...
02:26Protein Networks
An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
02:04Protein-protein Interfaces
Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a polypeptide...
01:24Protein Organization
Proteins are polymers of amino acid residues. They are versatile and responsible for different cellular functions, including DNA replication, molecular transport, catalysis, and structural support. Proteins have a hierarchical structure comprising at least three levels of organization: primary, secondary, and tertiary structure. Some large proteins have a quaternary structure where individual protein subunits are linked together.
The primary structure of a protein is its amino acid sequence.
The primary structure of a protein is its amino acid sequence.
00:49Ligand 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...
01:24Tagging and Fusion Proteins
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...