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submitted on 29.09.2020 and posted on 30.09.2020by Tomoki Himiyama, Yuko Tsuchiya, Yasushige Yonezawa, Tsutomu Nakamura
Direct control of protein quaternary structure (QS) is challenging owing
to the complexity of protein structure. As a protein with a characteristic QS,
peroxiredoxin from Aeropyrum pernix K1 (ApPrx) forms a decamer, wherein
five dimers associate to form a ring. Here, we disrupted and reconstituted
ApPrx QS via amino acid mutations and chemical modifications targeting hot
spots for protein assembly. The decameric QS of an ApPrx* mutant, wherein all
cysteine residues in wild-type ApPrx were mutated to serine, was destructed to
dimers via an F80C mutation. The dimeric ApPrx*F80C mutant was then modified
with a small molecule and successfully assembled as a decamer. Structural
analysis confirmed that an artificially installed chemical moiety potentially
facilitates suitable protein-protein interactions to rebuild a native
structure. Rebuilding of dodecamer was also achieved through an additional
amino acid mutation. This study describes a facile method to regulate protein