RESUMO
Although noncanonical amino acids (ncAAs) were first incorporated into phage libraries through amber suppression nearly two decades ago, their application for use in drug discovery has been limited due to inherent library bias towards sense-containing phages. Here, we report a technique based on superinfection immunity of phages to enrich amber-containing clones, thus avoiding the observed bias that has hindered incorporation of ncAAs into phage libraries. We then take advantage of this technique for development of active site-directed ligand evolution of peptides, where the ncAA serves as an anchor to direct the binding of its peptides to the target's active site. To demonstrate this, phage-displayed peptide libraries are developed that contain a genetically encoded butyryl lysine and are subsequently used to select for ligands that bind SIRT2. These ligands are then modified to develop low nanomolar inhibitors of SIRT2.
Assuntos
Âmbar/metabolismo , Bacteriófagos/metabolismo , Domínio Catalítico , Peptídeos/metabolismo , Descoberta de Drogas , Técnicas Genéticas , Humanos , Ligantes , Lisina/metabolismo , Simulação de Acoplamento Molecular , Biblioteca de Peptídeos , Sirtuína 2/metabolismoRESUMO
David Grimaldi introduces amber and the fossils contained therein.