TY - JOUR
T1 - Development of an Activity-Based Probe for Autotaxin
AU - Cavalli, Silvia
AU - Houben, Anna J.S.
AU - Albers, Harald M.H.G.
AU - Van Tilburg, Erica W.
AU - De Ru, Arnoud
AU - Aoki, Junken
AU - Van Veelen, Peter
AU - Moolenaar, Wouter H.
AU - Ovaa, Huib
PY - 2010/11/2
Y1 - 2010/11/2
N2 - 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.Marking biomarkers: ATX is a secreted lysophospholipase D that produces the lipid mediator lysophosphatidic acid. We have developed a fluorescent activity-based probe that covalently binds to the active site of ATX, allowing visualisation of active ATX. This probe can be used for monitoring ATX activity in body fluids and for inhibitor screening.
AB - 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.Marking biomarkers: ATX is a secreted lysophospholipase D that produces the lipid mediator lysophosphatidic acid. We have developed a fluorescent activity-based probe that covalently binds to the active site of ATX, allowing visualisation of active ATX. This probe can be used for monitoring ATX activity in body fluids and for inhibitor screening.
KW - Autotaxin
KW - Biomarkers
KW - Enzymes
KW - Inhibitors
KW - Lysophospholipase D
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U2 - 10.1002/cbic.201000349
DO - 10.1002/cbic.201000349
M3 - Article
C2 - 20941725
AN - SCOPUS:78049299270
VL - 11
SP - 2311
EP - 2317
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 16
ER -