Back to Search
Start Over
Unimodular Methylation by Adenylation–Thiolation Domains Containing an Embedded Methyltransferase.
- Source :
-
Journal of Molecular Biology . Oct2020, Vol. 432 Issue 21, p5802-5808. 7p. - Publication Year :
- 2020
-
Abstract
- Nonribosomal peptides (NRPs) are natural products that are biosynthesized by large multi-enzyme assembly lines called nonribosomal peptide synthetases (NRPSs). We have previously discovered that backbone or side chain methylation of NRP residues is carried out by an interrupted adenylation (A) domain that contains an internal methyltransferase (M) domain, while maintaining a monolithic AMA fold of the bifunctional enzyme. A key question that has remained unanswered is at which step of the assembly line mechanism the methylation by these embedded M domains takes place. Does the M domain methylate an amino acid residue tethered to a thiolation (T) domain on same NRPS module (in cis), or does it methylate this residue on a nascent peptide tethered to a T domain on another module (in trans)? In this study, we investigated the kinetics of methylation by wild-type AMAT tridomains from two NRPSs involved in biosynthesis of anticancer depsipeptides thiocoraline and echinomycin, and by mutants of these domains, for which methylation can occur only in trans. The analysis of the methylation kinetics unequivocally demonstrated that the wild-type AMATs methylate overwhelmingly in cis , strongly suggesting that this is also the case in the context of the entire NRPS assembly line process. The mechanistic insight gained in this study will facilitate rational genetic engineering of NRPS to generate unnaturally methylated NRPs. Unlabelled Image • Methylation mechanism in nonribosomal peptide synthetases remains to be determined. • Kinetic studies unequivocally established methylation occurs within one module. • This mechanistic logic will aid in engineering unnaturally methylated peptides. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00222836
- Volume :
- 432
- Issue :
- 21
- Database :
- Academic Search Index
- Journal :
- Journal of Molecular Biology
- Publication Type :
- Academic Journal
- Accession number :
- 146481692
- Full Text :
- https://doi.org/10.1016/j.jmb.2020.09.004