Premium
Enzyme‐Cleavable Linkers for Protein Chemical Synthesis through Solid‐Phase Ligations
Author(s) -
Abboud Skander A.,
Amoura Mehdi,
Madinier JeanBaptiste,
Renoux Brigitte,
Papot Sébastien,
Piller Véronique,
Aucagne Vincent
Publication year - 2021
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.202103768
Subject(s) - linker , chemistry , native chemical ligation , solid phase synthesis , combinatorial chemistry , cleavage (geology) , chemical ligation , peptide , enzyme , peptide synthesis , chemical synthesis , ligation , chemical modification , yield (engineering) , biochemistry , stereochemistry , in vitro , materials science , biology , microbiology and biotechnology , fracture (geology) , computer science , metallurgy , composite material , operating system
The total synthesis of long proteins requires the assembly of multiple fragments through successive ligations. The need for intermediate purification steps is a strong limitation, particularly in terms of overall yield. One solution to this problem would be solid‐supported chemical ligation (SPCL), for which a first peptide segment must be immobilized on a SPCL‐compatible solid support through a linker that can be cleaved under very mild conditions to release the assembled protein. The cleavage of SPCL linkers has previously required chemical conditions sometimes incompatible with sensitive protein targets. Herein, we describe an alternative enzymatic approach to trigger cleavage under extremely mild and selective conditions. Optimization of the linker structure and use of a small enzyme able to diffuse into the solid support were key to the success of the strategy. We demonstrated its utility by the assembly of three peptide segments on the basis of native chemical ligation to afford a 15 kDa polypeptide.