Biomaterial Database

Efficient incorporation of p-azido-l-phenylalanine into the protein using organic solvents. 2022

Xiaofeng Ma, and Bing Wei, and Enlin Wang

Department of Gynecology and Obstetrics, The Second Hospital of Anhui Medical University, Hefei, Anhui, 230601, China.

Azide, the most used photo-crosslinking group, facilitates the analysis of protein structure and function. This group is particularly useful when photochemically label antibodies and examine protein-protein interactions. The use of the expanded genetic code technique allows the special labeling of the functional azide group in proteins by adding the unnatural amino acid (UAA), p-azido-l-phenylalanine (AzF), in response to the amber codon during translation. However, a low UAA uptake rate due to mass transfer resistance in the cell membrane may lead to the early termination of the full-length protein. This study reports a general method for the efficient in vivo incorporation of AzF into the target protein by improving cell permeability using organic solvents. As expected, the yield of the full-length protein was significantly increased, which indicated that the AzF uptake was greatly improved due to the addition of organic solvents. Our method can serve as a good reference for improving the genetic incorporation of other kinds of UAAs into proteins.

UI	MeSH Term	Description	Entries
D010649	Phenylalanine	An essential aromatic amino acid that is a precursor of MELANIN; DOPAMINE; noradrenalin (NOREPINEPHRINE), and THYROXINE.	Endorphenyl,L-Phenylalanine,Phenylalanine, L-Isomer,L-Isomer Phenylalanine,Phenylalanine, L Isomer
D000596	Amino Acids	Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.	Amino Acid,Acid, Amino,Acids, Amino
D001386	Azides	Organic or inorganic compounds that contain the -N3 group.	Azide
D012997	Solvents	Liquids that dissolve other substances (solutes), generally solids, without any change in chemical composition, as, water containing sugar. (Grant & Hackh's Chemical Dictionary, 5th ed)	Solvent
D018388	Codon, Terminator	Any codon that signals the termination of genetic translation (TRANSLATION, GENETIC). PEPTIDE TERMINATION FACTORS bind to the stop codon and trigger the hydrolysis of the aminoacyl bond connecting the completed polypeptide to the tRNA. Terminator codons do not specify amino acids.	Amber Stop Codon,Codon, Amber Stop,Codon, Ochre Stop,Codon, Opal Stop,Codon, Stop,Stop Codon,Terminator Codon,Amber Codon,Amber Terminator Codon,Codon, Termination,Ochre Codon,Ochre Stop Codon,Opal Codon,Opal Stop Codon,Stop Codon UAA,Stop Codon UAG,Stop Codon UGA,Stop Signal, Translation,TAA Codon,TAG Codon,TGA Codon,UAA Codon,UAA Stop Codon,UAG Codon,UAG Stop Codon,UGA Codon,UGA Stop Codon,Amber Codons,Amber Stop Codons,Amber Terminator Codons,Codon UAA, Stop,Codon UAG, Stop,Codon UGA, Stop,Codon, Amber,Codon, Amber Terminator,Codon, Ochre,Codon, Opal,Codon, TAA,Codon, TAG,Codon, TGA,Codon, UAA,Codon, UAA Stop,Codon, UAG,Codon, UAG Stop,Codon, UGA,Codon, UGA Stop,Codons, Amber,Codons, Amber Stop,Codons, Amber Terminator,Codons, Ochre,Codons, Ochre Stop,Codons, Opal,Codons, Opal Stop,Codons, Stop,Codons, TAA,Codons, TAG,Codons, TGA,Codons, Termination,Codons, Terminator,Codons, UAA,Codons, UAA Stop,Codons, UAG,Codons, UAG Stop,Codons, UGA,Codons, UGA Stop,Ochre Codons,Ochre Stop Codons,Opal Codons,Opal Stop Codons,Stop Codon UGAs,Stop Codon, Amber,Stop Codon, Ochre,Stop Codon, Opal,Stop Codon, UAA,Stop Codon, UAG,Stop Codon, UGA,Stop Codons,Stop Codons, Amber,Stop Codons, Ochre,Stop Codons, Opal,Stop Codons, UAA,Stop Codons, UAG,Stop Codons, UGA,Stop Signals, Translation,TAA Codons,TAG Codons,TGA Codons,Termination Codon,Termination Codons,Terminator Codon, Amber,Terminator Codons,Terminator Codons, Amber,Translation Stop Signal,Translation Stop Signals,UAA Codons,UAA Stop Codons,UAA, Stop Codon,UAG Codons,UAG Stop Codons,UAG, Stop Codon,UGA Codons,UGA Stop Codons,UGA, Stop Codon