Site-specific dual encoding and labeling of proteins via genetic code expansion
Riley M Bednar1, P Andrew Karplus1, Ryan A Mehl1
1Department of Biochemistry and Biophysics, Oregon State University, 2011 Agricultural and Life Sciences Building, Corvallis, OR 97331-7305, USA; GCE4All Research Center, Oregon State University, 2011 Agricultural and Life Sciences, Corvallis, OR 97331-7305, USA.
Cell Chemical Biology
|March 28, 2023
Summary
Genetic code expansion (GCE) enables precise protein modification via dual encoding and labeling (DEAL). This powerful chemical biology tool allows site-specific incorporation of non-canonical amino acids for advanced biological engineering and study.
Area of Science:
- Chemical Biology
- Molecular Biology
- Biotechnology
Background:
- Selective protein modification at multiple sites is crucial for manipulating and studying living systems.
- Genetic code expansion (GCE) offers a method for site-specific protein modification with minimal functional disruption.
- The dual encoding and labeling (DEAL) process is a two-step approach within GCE for advanced protein engineering.
Purpose of the Study:
- To review the current state of dual encoding and labeling (DEAL) methodologies utilizing genetic code expansion (GCE).
- To elucidate the fundamental principles of GCE-based DEAL.
- To explore existing and potential applications of DEAL in biological research and engineering.
Main Methods:
- Summarizing the principles of GCE-based DEAL.
- Cataloging compatible encoding systems and bioorthogonal reactions.
- Reviewing demonstrated and potential applications of DEAL.
Main Results:
- The review details the foundational concepts of GCE-based DEAL.
- A comprehensive catalog of compatible GCE systems and labeling reactions is presented.
- Various applications and emerging paradigms in DEAL methodologies are explored.
Conclusions:
- GCE-based DEAL is a powerful chemical biology strategy for precise protein engineering.
- The review highlights current limitations and proposes future directions for DEAL methodologies.
- This approach holds significant promise for advancing the manipulation, engineering, and study of biological systems.
Keywords:
DEALbioorthogonal chemistrydual encoding and labelinggenetic code expansionprotein bioconjugationprotein engineeringMore Related Videos
11:47Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System
Published on: August 1, 2016
16.0K
13:11Super-Resolution Imaging of Bacterial Secreted Proteins Using Genetic Code Expansion
Published on: February 10, 2023
1.5K
Related Concept Videos
03:06From DNA to Protein
18.7K
The flow of genetic information in cells from DNA to mRNA to protein is described by the central dogma, which states that genes specify the sequence of mRNAs, which in turn specify the sequence of amino acids making up all proteins. The decoding of one molecule to another is performed by specific proteins and RNAs. Because the information stored in DNA is so central to cellular function, it makes intuitive sense that the cell would make mRNA copies of this information for protein synthesis...
18.7K
01:24Tagging and Fusion Proteins
6.8K
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...
6.8K
01:25The Central Dogma
127.3K
Overview
127.3K
02:28Leaky Scanning
5.2K
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.2K
02:24Ribosome Profiling
3.6K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.6K
02:26tRNA Activation
19.5K
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
19.5K