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Bioorthogonal Ligation Using Site‐Specific Tetrazine‐Alkene Coupling On Proteins
Author(s) -
Refakis Christian André,
Blackman Melissa L,
Fox Joseph M,
Mehl Ryan A
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.912.7
Subject(s) - bioorthogonal chemistry , alkene , tetrazine , chemistry , chemical ligation , combinatorial chemistry , covalent bond , amino acid , bioconjugation , native chemical ligation , chemical biology , reactivity (psychology) , intein , biochemistry , chemical synthesis , click chemistry , organic chemistry , in vitro , catalysis , medicine , rna , alternative medicine , pathology , rna splicing , gene
The selective coupling of chemical labels to proteins is key to many applications in medicine and biotechnology. Chemical labeling strategies require two functional groups to react rapidly and selectively with one another under biological conditions to form a stable covalent bond. Despite their great utility, few effective bioorthogonal ligation reactions exist that are non‐toxic and react specifically to the programmed functionality. The objective of this study is to expand on the array of available bioorthogonal ligation reactions by using site‐specific unnatural amino acid incorporation to exploit the alkene‐tetrazine reaction. The amino acid p‐vinyl‐L‐phenylalanine (pVF) was synthesized and incorporated into proteins site‐specifically, in vivo, using an unnatural aminoacyl‐tRNA synthetase (RS) pair. The reactivity of a diverse family of tetrazines was explored with pVF‐protein. ESI‐MS confirmed the tetrzine‐alkene ligation reaction occurs with high fidelity, site‐specificity, and yield. We expect the results of this work will provide scientists with an additional venue for attaching small chemical labels and/or drugs to proteins.