TitleGenetically encoded tetrazine amino acid directs rapid site-specific in vivo bioorthogonal ligation with trans-cyclooctenes.
Publication TypeJournal Article
Year of Publication2012
AuthorsSeitchik JL, Peeler JC, Taylor MT, Blackman ML, Rhoads TW, Cooley RB, Refakis C, Fox JM, Mehl RA
JournalJ Am Chem Soc
Date Published2012 Feb 15
KeywordsAmino Acids, Catalysis, Chemistry, Cyclooctanes, Escherichia coli, Genetic Engineering, Green Fluorescent Proteins, Methanococcus, Models, Chemical, Molecular Conformation, Proteins, Pyridines, Recombinant Proteins, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Tyrosine-tRNA Ligase

Bioorthogonal ligation methods with improved reaction rates and less obtrusive components are needed for site-specifically labeling proteins without catalysts. Currently no general method exists for in vivo site-specific labeling of proteins that combines fast reaction rate with stable, nontoxic, and chemoselective reagents. To overcome these limitations, we have developed a tetrazine-containing amino acid, 1, that is stable inside living cells. We have site-specifically genetically encoded this unique amino acid in response to an amber codon allowing a single 1 to be placed at any location in a protein. We have demonstrated that protein containing 1 can be ligated to a conformationally strained trans-cyclooctene in vitro and in vivo with reaction rates significantly faster than most commonly used labeling methods.

Alternate JournalJ. Am. Chem. Soc.
PubMed ID22283158
PubMed Central IDPMC3369569
Grant ListP20 RR017716 / RR / NCRR NIH HHS / United States
P30ES000210 / ES / NIEHS NIH HHS / United States
/ / Howard Hughes Medical Institute / United States