Bypassing the requirement for aminoacyl-tRNA by a cyclodipeptide synthase enzyme.
| Autor: | Harding CJ; School of Biology, Biomedical Sciences Research Complex, University of St Andrews, St Andrews Fife KY16 9ST UK cmc27@st-andrews.ac.uk., Sutherland E; School of Biology, Biomedical Sciences Research Complex, University of St Andrews, St Andrews Fife KY16 9ST UK cmc27@st-andrews.ac.uk., Hanna JG; Arab Academy for Science, Technology, and Maritime Transport (AASTMT) Cairo Campus Egypt., Houston DR; Institute of Quantitative Biology, Biochemistry and Biotechnology, University of Edinburgh Waddington 1 Building, King's Buildings Edinburgh EH9 3BF UK., Czekster CM; School of Biology, Biomedical Sciences Research Complex, University of St Andrews, St Andrews Fife KY16 9ST UK cmc27@st-andrews.ac.uk. |
|---|---|
| Jazyk: | angličtina |
| Zdroj: | RSC chemical biology [RSC Chem Biol] 2021 Jan 15; Vol. 2 (1), pp. 230-240. Date of Electronic Publication: 2021 Jan 15. |
| DOI: | 10.1039/d0cb00142b |
| Abstrakt: | Cyclodipeptide synthases (CDPSs) produce a variety of cyclic dipeptide products by utilising two aminoacylated tRNA substrates. We sought to investigate the minimal requirements for substrate usage in this class of enzymes as the relationship between CDPSs and their substrates remains elusive. Here, we investigated the Bacillus thermoamylovorans enzyme, BtCDPS, which synthesises cyclo(l-Leu-l-Leu). We systematically tested where specificity arises and, in the process, uncovered small molecules (activated amino esters) that will suffice as substrates, although catalytically poor. We solved the structure of BtCDPS to 1.7 Å and combining crystallography, enzymatic assays and substrate docking experiments propose a model for how the minimal substrates interact with the enzyme. This work is the first report of a CDPS enzyme utilizing a molecule other than aa-tRNA as a substrate; providing insights into substrate requirements and setting the stage for the design of improved simpler substrates. Competing Interests: There are no conflicts to declare. (This journal is © The Royal Society of Chemistry.) |
| Databáze: | MEDLINE |
| Externí odkaz: |
|