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

Christian KappeZainelabdeen H MohamedEyad NaserAlexander CarpinteiroChristoph Arenz
Published in: Chemistry (Weinheim an der Bergstrasse, Germany) (2020)
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.
Keyphrases
  • living cells
  • fluorescent probe
  • single molecule
  • flow cytometry
  • acute kidney injury
  • lipopolysaccharide induced
  • dna binding
  • energy transfer
  • transcription factor
  • aqueous solution