Recognition-induced covalent capturing and labeling as a general strategy for protein detection

In this work we have developed a peptide-based method for protein detection, termed as “Recognition-induced Covalent Capturing and Labeling” (RCCL). In this method, upon binding of the peptide with the target protein, electrochemically controlled and metal catalyzed oxidative cross-linking can be induced between the peptide and the target protein. Specifically, the peptide and the target protein are cross-linked by the formation of dityrosine between tyrosine moieties of the two molecules. Meanwhile, the dityrosine formed in this manner also has fluorescent signal readout. Therefore, the proposed method needs only one probe for the target protein, and the initial non-covalent molecular recognition can be finalized by cross-linking between the peptide and the target, while the dityrosine formed between peptide and protein can also act as a signal reporter, thereby greatly simplifying the design. Moreover, the robust covalent capturing via RCCL also enables detection in complex biological and clinical samples. These results point to the prospect of using RCCL as a promising method in protein detection in the future.