Application of feature-based molecular networking and MassQL for the MS/MS fragmentation study of depsipeptides

Abstract

The Feature-based Molecular Networking (FBMN) is a well-known approach for mapping and identifying structures and analogues. However, in the absence of prior knowledge about the molecular class, assessing specific fragments and clusters requires time-consuming manual validation. This study demonstrates that combining FBMN and Mass Spec Query Language (MassQL) is an effective strategy for accelerating the decoding mass fragmentation pathways and identifying molecules with comparable fragmentation patterns, such as beauvericin and its analogues. To accomplish this objective, a spectral similarity network was built from ESI-MS/MS experiments of Fusarium oxysporum at various collision energies (CIDs) and paired with a MassQL search query for conserved beauvericin ions. FBMN analysis revealed that sodiated and protonated ions clustered differently, with sodiated adducts needing more collision energy and exhibiting a distinct fragmentation pattern. Based on this distinction, two sets of particular fragments were discovered for the identification of these hexadepsipeptides: ([M + H]+) m/z 134, 244, 262, and 362 and ([M + Na]+) m/z 266, 284 and 384. By using these fragments, MassQL accurately found other analogues of the same molecular class and annotated beauvericins that were not classified by FBMN alone. Furthermore, FBMN analysis of sodiated beauvericins at 70 eV revealed subclasses with distinct amino acid residues, allowing distinction between beauvericins (beauvericin and beauvericin D) and two previously unknown structural isomers with an unusual methionine sulfoxide residue. In summary, our integrated method revealed correlations between adduct types and fragmentation patterns, facilitated the detection of beauvericin clusters, including known and novel analogues, and allowed for the differentiation between structural isomers.

Comprehensive Mass Spectrometric Metabolomic Profiling of a Chemically Diverse Collection of Plants of the Celastraceae Family

“…It can be employed to develop chemotaxonomic models since it contains the quality controls and analytical replicates necessary (See Technical validation). Researchers can use the Mass Spec Query Language (MassQL) to interrogate the data set for specific spectral patterns produced by a certain group of compounds 62,63 . For example, this MassQL query searches for the MS 2 fragments of the 9-oxodihydro-β-agarofuran 60 mentioned above.…”

Integration of Wnt-inhibitory activity and structural novelty scoring results to uncover novel bioactive natural products: new Bicyclo[3.3.1]non-3-ene-2,9-diones from the leaves of Hymenocardia punctata

“…Early structural identifications of NPs in NEs can assist researchers in avoiding reported active NPs or efficiently searching for analogs of previously reported bioactive NPs ( Hubert et al, 2017 ; Selegato et al, 2023 ). With the advancement of computational annotations methods throughput in metabolomics, it is now possible to evaluate the chemical space of large NEs collections ( Gaudry et al, 2023 ).…”

Integration of Wnt-inhibitory Activity and Structural Novelty Scoring Results to Uncover Novel Bioactive Natural Products: New Bicyclo[3.3.1]non-3-ene-2,9-diones from the Leaves of Hymenocardia punctata.

“…Early structural identifications of NPs in NEs can assist researchers in avoiding reported active NPs or efficiently searching for analogs of previously reported bioactive NPs (Hubert et al, 2017;Selegato et al, 2023). With the advancement of computational annotations methods throughput in metabolomics, it is now possible to evaluate the chemical space of large NEs collections .…”