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Development of an Activity‐Based Probe for Autotaxin
Author(s) -
Cavalli Silvia,
Houben Anna J. S.,
Albers Harald M. H. G.,
van Tilburg Erica W.,
de Ru Arnoud,
Aoki Junken,
van Veelen Peter,
Moolenaar Wouter H.,
Ovaa Huib
Publication year - 2010
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201000349
Subject(s) - autotaxin , chemistry , computational biology , biochemistry , nanotechnology , biology , lysophosphatidic acid , materials science , receptor
Autotaxin (ATX), or ecto‐nucleotide pyrophosphatase/phosphodiesterase 2 (ENPP2), is a secreted lysophospholipase D that hydrolyses lysophosphatidylcholine into the lipid mediator lysophosphatidic acid (LPA), a mitogen and chemoattractant for many cell types. ATX has been implicated in tumour progression and inflammation, and might serve as a biomarker. Here we describe the development of a fluorescent activity‐based probe that covalently binds to the active site of ATX. The probe consists of a lysophospholipid‐based backbone linked to a trapping moiety that becomes reactive after phosphate ester hydrolysis, and a Cy5 fluorescent dye to allow visualisation of active ATX. The probe reacts specifically with the three known isoforms of ATX, it competes with small‐molecule inhibitors for binding to ATX and allows ATX activity in plasma to be determined. Our activity‐based reporter will be useful for monitoring ATX activity in biological fluids and for inhibitor screening.