Sortase-Catalyzed Protein Domain Inversion.
Angew Chem Int Ed Engl
; 63(14): e202316777, 2024 04 02.
Article
em En
| MEDLINE
| ID: mdl-38366985
ABSTRACT
Topological transformations and permutations of proteins have attracted significant interest as strategies to generate new protein functionalities or stability. These efforts have mainly been inspired by naturally occurring post-translational modifications, such as head-to-tail cyclization, circular permutation, or lasso-like entanglement. Such approaches can be realized experimentally via genetic encoding, in the case of circular permutation, or via enzymatic processing, in the case of cyclization. Notably, these previously described strategies leave the polypeptide backbone orientation unaltered. Here we describe an unnatural protein permutation, the protein domain inversion, whereby a C-terminal portion of a protein is enzymatically inverted from the canonical N-to-C to a C-to-C configuration with respect to the N-terminal part of the protein. The closest conceptually analogous biological process is perhaps the inversion of DNA segments as catalyzed by recombinases. We achieve these inversions using an engineered sortase A, a widely used transpeptidase. Our reactions proceed efficiently under mild conditions at 4-25 °C and are compatible with entirely heterologously-produced protein substrates.
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Base de dados:
MEDLINE
Assunto principal:
Peptidil Transferases
/
Aminoaciltransferases
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article