Title | Genetically encoded tetrazine amino acid directs rapid site-specific in vivo bioorthogonal ligation with trans-cyclooctenes. |
Publication Type | Journal Article |
Year of Publication | 2012 |
Authors | Seitchik JL, Peeler JC, Taylor MT, Blackman ML, Rhoads TW, Cooley RB, Refakis C, Fox JM, Mehl RA |
Journal | J Am Chem Soc |
Volume | 134 |
Issue | 6 |
Pagination | 2898-901 |
Date Published | 2012 Feb 15 |
ISSN | 1520-5126 |
Keywords | Amino 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 |
Abstract | 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. |
DOI | 10.1021/ja2109745 |
Alternate Journal | J. Am. Chem. Soc. |
PubMed ID | 22283158 |
PubMed Central ID | PMC3369569 |
Grant List | P20 RR017716 / RR / NCRR NIH HHS / United States P30ES000210 / ES / NIEHS NIH HHS / United States / / Howard Hughes Medical Institute / United States |