A Novel Visible Range FRET Probe for Monitoring Acid Sphingomyelinase Activity in Living Cells. Issue 26 (21st April 2020)

• Record Type: Journal Article
• Authors: Kappe, Christian; Mohamed, Zainelabdeen H.; Naser, Eyad; Carpinteiro, Alexander; Arenz, Christoph
• Is Part Of: Chemistry. Volume 26:Issue 26(2020)
• Page Start: 5780
• Page End: 5783
• Publication Date: 2020-04-21
• Subjects: enzyme assays -- flow cytometry -- Förster resonance energy transfer -- live cell assays -- sphingolipids; Chemistry -- Periodicals; 540
• DOI: 10.1002/chem.202000133
• Languages: English
• ISSNs: 0947-6539

Abstract

Abstract: Activity of acid sphingomyelinase has been implicated in a number of diseases like acute lung injury, sepsis or metastasis of melanoma cells. Here, we present a sphingomyelinase FRET probe based on FAM/BODIPY dyes for real‐time monitoring of acid sphingomyelinase. The probe gives rise to a tremendous increase in fluorescence of the fluorescein FRET donor upon cleavage and we show that this is, to a significant part, due to cleavage‐associated phase transition, suggesting a more systematic consideration of such effects for future probe development. The probe allows for the first time to monitor relative sphingomyelinase activities of intact living cells by flow cytometry. Abstract : More than just FRET : A visible range FRET probe for acid sphingomyelinase (ASM) was developed. The fluorescence increase is >100 fold and additionally fueled by environmental sensitivity of the fluorescein dye. The probe was used for analysis of live cell ASM activities by flow cytometry.