title: Reprogramming nonribosomal peptide synthetases for "clickable" amino acids.
abstract: Nonribosomal peptide synthetases (NRPSs) are multifunctional enzymes that produce a wide array of bioactive peptides. Here we show that a single tryptophan-to-serine mutation in phenylalanine-specific NRPS adenylation domains enables the efficient activation of non-natural aromatic amino acids functionalized with azide and alkyne groups. The resulting 10(5)-fold switch in substrate specificity was achieved without appreciable loss of catalytic efficiency. Moreover, the effective communication of the modified A domains with downstream modules in dipeptide synthetases permitted incorporation of O-propargyl-L-tyrosine into diketopiperazines both in vitro and in vivo, even in the presence of competing phenylalanine. Because azides and alkynes readily undergo bioorthogonal click reactions, reprogramming NRPSs to accept non-natural amino acids that contain these groups provides a potentially powerful means of isolating, labeling, and modifying biologically active peptides.
authors: Laboratory of Organic Chemistry, ETH Zürich, 8093 Zürich (Switzerland).
authors: Hajo
authors: H
authors: Kries
authors: Rudolf
authors: R
authors: Wachtel
authors: Anja
authors: A
authors: Pabst
authors: Benedikt
authors: B
authors: Wanner
authors: David
authors: D
authors: Niquille
authors: Donald
authors: D
authors: Hilvert
mesh_terms: D000480:Alkynes / Q000737:chemistry / Q000378:metabolism; D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism*; D001386:Azides / Q000737:chemistry / Q000378:metabolism; D057930:Click Chemistry; D008958:Models, Molecular; D015394:Molecular Structure; D010453:Peptide Synthases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000480:Alkynes; D000596:Amino Acids; D001386:Azides; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase
keywords: adenylation domain; click chemistry; diketopiperazine; mutagenesis; nonribosomal peptide