Using MbtH‐Like Proteins to Alter the Substrate Profile of a Nonribosomal Peptide Adenylation Enzyme

• Published in: Chembiochem : a European journal of chemical biology Vol. 19; no. 20; pp. 2186 - 2194
• Main Authors: Mori, Shogo, Green, Keith D., Choi, Ryan, Buchko, Garry W., Fried, Michael G., Garneau‐Tsodikova, Sylvie
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 18.10.2018; ChemPubSoc Europe
• Subjects: Adenylation; antibiotics, synthethic biology, non-ribosomal peptide synthetases, SSGCID; Bacteria - metabolism; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacterial Proteins - physiology; Bioinformatics; Biosynthesis; Domains; enzymes; Kinetics; Peptide Synthases - chemistry; Peptide Synthases - metabolism; Peptides; Protein Domains; Proteins; protein–protein interactions; Substrate Specificity; Substrates; enzymes; kinetics; bioinformatics; biosynthesis; protein-protein interactions
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.201800240

MbtH‐like proteins (MLPs) are required for soluble expression and/or optimal activity of some adenylation (A) domains of nonribosomal peptide synthetases. Because A domains can interact with noncognate MLP partners, how the function of an A domain, TioK, involved in the biosynthesis of the bisintercalator thiocoraline, is altered by noncognate MLPs has been investigated. Measuring TioK activity with 12 different MLPs from a variety of bacterial species by using a radiometric assay suggested that the A domain substrate promiscuity could be altered by foreign MLPs. Kinetic studies and bioinformatics analysis expanded the complexity of MLP functions and interactions. TioK profile picture: MbtH‐like proteins (MLPs) are required for the soluble expression and optimal activity of some adenylation (A) domains of nonribosomal peptide synthetases. A study of 11 noncognate MLPs from various nonribosomal peptide synthetase assembly lines expands the substrate profile of the TioK A domain from the thiocoraline biosynthetic gene cluster, in comparison with the cognate MLP TioT.