Apr 25, 2020

Computationally-guided exchange of substrate selectivity motifs in a modular polyketide synthase acyltransferase

BioRxiv : the Preprint Server for Biology
E. KalkreuterGavin John Williams

Abstract

Acyltransferases (ATs) of modular polyketide synthases catalyze the installation of malonyl-CoA extenders into polyketide scaffolds. Subsequently, AT domains have been targeted extensively to site-selectively introduce various extenders into polyketides. Yet, a complete inventory of AT residues responsible for substrate selection has not been established, critically limiting the efficiency and scope of AT engineering. Here, molecular dynamics simulations were used to prioritize ~50 mutations in the active site of EryAT6 from erythromycin biosynthesis. Following detailed in vitro studies, 13 mutations across 10 residues were identified to significantly impact extender unit selectivity, including nine residues that were previously unassociated with AT specificity. Unique insights gained from the MD studies and the novel EryAT6 mutations led to identification of two previously unexplored structural motifs within the AT active site. Remarkably, exchanging both motifs in EryAT6 with those from ATs with unusual extender specificities provided chimeric PKS modules with expanded and inverted substrate specificity. Our enhanced understanding of AT substrate selectivity and application of this motif-swapping strategy is expected to advan...Continue Reading

  • References
  • Citations

References

  • We're still populating references for this paper, please check back later.
  • References
  • Citations

Citations

  • This paper may not have been cited yet.

Mentioned in this Paper

Dysequilibrium Syndrome
Persons
Participant
Epidemiologic Studies
Simulation
Epidemiology
Analysis
Genetic Linkage
Silo (Dataset)
Genotype Determination

Related Feeds

BioRxiv & MedRxiv Preprints

BioRxiv and MedRxiv are the preprint servers for biology and health sciences respectively, operated by Cold Spring Harbor Laboratory. Here are the latest preprint articles (which are not peer-reviewed) from BioRxiv and MedRxiv.

Related Papers

Genetic Epidemiology
Timothy Shin Heng MakPak Chung Sham
American Journal of Human Genetics
Bjarni VilhjalmssonNick J Patterson
BioRxiv : the Preprint Server for Biology
Bjarni VilhjalmssonAlkes L Price
BioRxiv : the Preprint Server for Biology
Timothy Shin Heng MakPak Chung Sham
© 2020 Meta ULC. All rights reserved