Rational engineering of allosteric protein switches by in silico prediction of domain insertion sites.

ABSTRACT: Domain insertion engineering is a powerful approach to juxtapose otherwise separate biological functions, resulting in proteins with new-to-nature activities. A prominent example are switchable protein variants, created by receptor domain insertion into effector proteins. Identifying suitable, allosteric sites for domain insertion, however, typically requires extensive screening and optimization. We present ProDomino, a machine learning pipeline to rationalize domain recombination, trained on a semisynthetic protein sequence dataset derived from naturally occurring intradomain insertion events. ProDomino robustly identifies domain insertion sites in proteins of biotechnological relevance, which we experimentally validated in Escherichia coli and human cells. Finally, we used light- and che

SUBMITTER: Wolf B 

PROVIDER: S-EPMC12328240 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature