RESUMEN
A Natural Compound Library containing myxobacterial secondary metabolites was screened in murine macrophages for novel activators of IL-1ß maturation and secretion. The most potent of three hits in total was a so far undescribed metabolite, which was identified from the myxobacterium Hyalangium minutum strain Hym3. While the planar structure of 1 was elucidated by high resolution mass spectrometry and NMR data yielding an asymmetric boron containing a macrodiolide core structure, its relative stereochemistry of all 20 stereocenters of the 42-membered ring was assigned by rotating frame Overhause effect spectroscopy correlations, 1H,1H, and 1H,13C coupling constants, and by comparison of 13C chemical shifts to those of the structurally related metabolites tartrolon B-D. The absolute stereochemistry was subsequently assigned by Mosher's and Marfey's methods. Further functional studies revealed that hyaboron and other boronated natural compounds resulted in NLRP3 inflammasome dependent IL-1ß maturation, which is most likely due to their ability to act as potassium ionophores. Moreover, besides its inflammasome-stimulatory activity in human and mouse cells, hyaboron (1) showed additional diverse biological activities, including antibacterial and antiparasitic effects.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Compuestos de Boro/farmacología , Macrólidos/farmacología , Myxococcales/química , Adyuvantes Inmunológicos/química , Animales , Antibacterianos/química , Antibacterianos/farmacología , Antifúngicos/química , Antifúngicos/farmacología , Antineoplásicos/química , Antineoplásicos/farmacología , Compuestos de Boro/química , Línea Celular Tumoral , Hongos/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Humanos , Inflamasomas/metabolismo , Macrólidos/química , Ratones , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Bibliotecas de Moléculas Pequeñas/química , EstereoisomerismoRESUMEN
The flavonol branch of flavonoid biosynthesis is under transcriptional control of the R2R3-MYBs production of flavonol glycoside1 (PFG1/MYB12, PFG2/MYB11 and PFG3/MYB111) in Arabidopsis thaliana. Here, we investigated the influence of specific PFG transcription factors on flavonol distribution in various organs. A combination of genetic and metabolite analysis was used to identify transcription factor gene-metabolite correlations of the flavonol metabolic pathway. Flavonol glycoside accumulation patterns have been analysed in wild-type and multiple R2R3-MYB PFG mutants in an organ- and development-dependent manner using high-performance thin-layer chromatography, supplemented with liquid chromatography-mass spectroscopy metabolite profiling. Our results clearly demonstrate a differential influence of MYB11, MYB12 and MYB111 on the spatial accumulation of specific flavonol derivatives in leaves, stems, inflorescences, siliques and roots. In addition, MYB11-, MYB12- and MYB111-independent flavonol glycoside accumulation was observed in pollen grains and siliques/seeds. The highly complex tissue- and developmental-specific regulation of flavonol biosynthesis in A. thaliana is orchestrated by at least four PFG transcription factors, differentially influencing the spatial accumulation of specific flavonol derivatives. We present evidence that a separate flavonol control mechanism might be at play in pollen.