Development of a novel FRET probe for the real-time determination of ceramidase activity

Krishna P Bhabak; Anett Hauser; Susanne Redmer; Sebastian Banhart; Dagmar Heuer; Christoph Arenz

doi:10.1002/cbic.201300207

Development of a novel FRET probe for the real-time determination of ceramidase activity

Chembiochem. 2013 Jun 17;14(9):1049-52. doi: 10.1002/cbic.201300207. Epub 2013 May 31.

Authors

Krishna P Bhabak¹, Anett Hauser, Susanne Redmer, Sebastian Banhart, Dagmar Heuer, Christoph Arenz

Affiliation

¹ Department of Organic & Bioorganic Chemistry, Humboldt Universität zu Berlin, Institute for Chemistry, Brook-Taylor Str-2, 12489 Berlin, Germany.

PMID: 23728798
DOI: 10.1002/cbic.201300207

Abstract

Fretful novelty: We developed two novel doubly labelled fluorescent ceramide analogues that exhibit significant FRET and undergo hydrolysis by ceramidases. We present a fluorescent sphingolipid FRET probe that allows homogeneous ratiometric determination of enzyme activity in real-time.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Ceramidases / metabolism*
Ceramides / chemistry*
Ceramides / metabolism
Fluorescence Resonance Energy Transfer*
Fluorescent Dyes / chemistry*
Golgi Apparatus / metabolism
HeLa Cells
Humans
Hydrolysis
Microscopy, Fluorescence
Oxadiazoles / chemistry
Oxazines / chemistry
Phospholipases A2 / metabolism

Substances

7-nitrobenz-2-oxa-1,3-diazole
Ceramides
Fluorescent Dyes
Oxadiazoles
Oxazines
Phospholipases A2
Ceramidases
nile red