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Abstract Here we report the design ofN²‐carboxy‐4‐aryl‐1,2,3‐triazole‐lysines (CATKs) and their site‐specific incorporation into proteins via genetic code expansion. When introduced into the protein dimer interface, CATKs permitted spontaneous, proximity‐driven, site‐selective crosslinking to generate covalent protein dimers in living cells, with phenyl‐bearing CATK‐1exhibiting high reactivity toward the proximal Lys and Tyr. Furthermore, when introduced into theN‐terminal β‐strand of either a single‐chain VHH antibody or a supercharged monobody, CATK‐1enabled site‐specific, inter‐strand, orthogonal crosslinking with a proximal Tyr located on the opposing β‐strand. Compared with a non‐crosslinked monobody, the orthogonally crosslinked monobody displayed improved cellular uptake and enhanced proteolytic stability against an endosomal enzyme. The robust crosslinking reactivity of CATKs should facilitate the design of novel protein topologies with improved physicochemical properties.