Project description:For peloid natural product discovery we have produced 12 spectral libraries, 6 for nighttime sampled peloid (.mgf libraries containing suffix "N") and 6 for daytime sampled peloid (.mgf libraries containing suffix "D" in their name), containing a total of 14,417 individual mass spectra, using a Quadrupole Time-of-Flight (qTOF; 6546 LC/Q-TOF, Agilent, USA) LC-HRMS. In order to identify known natural products, preferably with predetermined biological activity, which would enable us to evaluate Nin peloid mud medicinal property claims, we have used freely available COlleCtion of Open Natural prodUcTs (COCONUT).
Project description:Finding the targets of natural products is of key importance in both chemical biology and drug discovery, and deconvolution of cofactor interactomes contribute to the functional annotation of the proteome. Identifying the proteins that underlie natural compound activity in phenotypic screens help to validate the respective targets and, potentially, expand the druggable proteome. Here, we present a generally applicable protocol for the photoactivated immobilisation of unmodified and microgram quantities of natural products on diazirine-decorated beads and their use for systematic affinity-based proteome profiling. We show that amongst 31 molecules of very diverse reported activity and biosynthetic origin, 25 could indeed be immobilised. Dose-response competition binding experiments using lysates of human or bacterial cells followed by quantitative mass spectrometry recapitulated <100 µM targets of 9 molecules. Among them, immobilisation of coenzyme A produced a tool to interrogate proteins containing a HotDog domain. Surprisingly, immobilisation of the cofactor flavin adenine dinucleotide (FAD) led to the identification of nanomolar interactions with dozens of RNA-binding proteins.
2022-10-06 | PXD033292 | Pride
Project description:Effect of Natural Products on the Intestinal Microbial