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A Novel Visible Range FRET Probe for Monitoring Acid Sphingomyelinase Activity in Living Cells.

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.

SUBMITTER: Kappe C

PROVIDER: S-EPMC7317515 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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A Novel Visible Range FRET Probe for Monitoring Acid Sphingomyelinase Activity in Living Cells.

Kappe Christian C Mohamed Zainelabdeen H ZH Naser Eyad E Carpinteiro Alexander A Arenz Christoph C

Chemistry (Weinheim an der Bergstrasse, Germany) 20200421 26

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 consid ...[more]

PMID: 32092185

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