Native Metabolomics Identifies the Rivulariapeptolide Family of Protease Inhibitors

Reher, R.; Aron, A. T.; Fajtova, P.; Liu, C.; Ben Shalom, I. Y.; Bittremieux, W.; Wang, M.; Matos-Hernandez, M. L.; Alexander, K. L.; Caro-Diaz, E. J.; Naman, C. B.; Dorrestein, P. C.; O'Donoghue, A. J.; Gerwick, W. H.; Petras, D.

doi:10.1101/2021.09.03.458897

bioRxiv · 10.1101/2021.09.03.458897

Native Metabolomics Identifies the Rivulariapeptolide Family of Protease Inhibitors

Reher, R.·Aron, A. T.·Fajtova, P.·Liu, C.·Ben Shalom, I. Y.·Bittremieux, W.·Wang, M.·Matos-Hernandez, M. L.·Alexander, K. L.·Caro-Diaz, E. J.·Naman, C. B.·Dorrestein, P. C.·O'Donoghue, A. J.·Gerwick, W. H.·Petras, D.

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Abstract

The identity and biological activity of most metabolites still remain unknown. A key bottleneck in the full exploration of this tremendous source of new structures and pharmaceutical activities is the compound purification needed for bioactivity assignments of individual compounds and downstream structure elucidation. To enable bioactivity-focused compound identification from complex mixtures, we developed a scalable native metabolomics approach that integrates non-targeted liquid chromatography tandem mass spectrometry, and simultaneous detection of protein binding via native mass spectrometry. While screening for new protease inhibitors from an environmental cyanobacteria community, native metabolomics revealed 30 cyclodepsipeptides as chymotrypsin binders. Mass spectrometry-guided purification then allowed for the full structure elucidation of four new specialized metabolites via tandem mass spectrometry, chemical derivatization, and nuclear magnetic resonance spectroscopy. Together with the evaluation of biological activities, our results identified the rivulariapeptolides as a family of serine protease inhibitors with nanomolar potency, highlighting native metabolomics as promising approach for drug discovery, chemical ecology, and chemical biology studies.

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Reher, R., Aron, A. T., Fajtova, P., Liu, C., Ben Shalom, I. Y., Bittremieux, W., Wang, M., Matos-Hernandez, M. L., Alexander, K. L., Caro-Diaz, E. J., Naman, C. B., Dorrestein, P. C., O'Donoghue, A. J., Gerwick, W. H., Petras, D.. 2021-09-04. Native Metabolomics Identifies the Rivulariapeptolide Family of Protease Inhibitors. https://doi.org/10.1101/2021.09.03.458897

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