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Olaparib‐Based Photoaffinity Probes for PARP‐1 Detection in Living Cells

The poly‐ADP‐ribose polymerase (PARP) is a protein from the family of ADP‐ribosyltransferases that catalyzes polyadenosine diphosphate ribose (ADPR) formation in order to attract the DNA repair machinery to sites of DNA damage. The inhibition of PARP activity by olaparib can cause cell death, which...

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Autores principales: Voorneveld, Jim, Florea, Bogdan I., Bakkum, Thomas, Mendowicz, Rafal J., van der Veer, Miriam S., Gagestein, Berend, van Kasteren, Sander I., van der Stelt, Mario, Overkleeft, Herman S., Filippov, Dmitri V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7496120/
https://www.ncbi.nlm.nih.gov/pubmed/32282108
http://dx.doi.org/10.1002/cbic.202000042
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author Voorneveld, Jim
Florea, Bogdan I.
Bakkum, Thomas
Mendowicz, Rafal J.
van der Veer, Miriam S.
Gagestein, Berend
van Kasteren, Sander I.
van der Stelt, Mario
Overkleeft, Herman S.
Filippov, Dmitri V.
author_facet Voorneveld, Jim
Florea, Bogdan I.
Bakkum, Thomas
Mendowicz, Rafal J.
van der Veer, Miriam S.
Gagestein, Berend
van Kasteren, Sander I.
van der Stelt, Mario
Overkleeft, Herman S.
Filippov, Dmitri V.
author_sort Voorneveld, Jim
collection PubMed
description The poly‐ADP‐ribose polymerase (PARP) is a protein from the family of ADP‐ribosyltransferases that catalyzes polyadenosine diphosphate ribose (ADPR) formation in order to attract the DNA repair machinery to sites of DNA damage. The inhibition of PARP activity by olaparib can cause cell death, which is of clinical relevance in some tumor types. This demonstrates that quantification of PARP activity in the context of living cells is of great importance. In this work, we present the design, synthesis and biological evaluation of photo‐activatable affinity probes inspired by the olaparib molecule that are equipped with a diazirine for covalent attachment upon activation by UV light and a ligation handle for the addition of a reporter group of choice. SDS‐PAGE, western blotting and label‐free LC‐MS/MS quantification analysis show that the probes target the PARP‐1 protein and are selectively outcompeted by olaparib; this suggests that they bind in the same enzymatic pocket. Proteomics data are available via ProteomeXchange with identifier PXD018661.
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spelling pubmed-74961202020-09-25 Olaparib‐Based Photoaffinity Probes for PARP‐1 Detection in Living Cells Voorneveld, Jim Florea, Bogdan I. Bakkum, Thomas Mendowicz, Rafal J. van der Veer, Miriam S. Gagestein, Berend van Kasteren, Sander I. van der Stelt, Mario Overkleeft, Herman S. Filippov, Dmitri V. Chembiochem Communications The poly‐ADP‐ribose polymerase (PARP) is a protein from the family of ADP‐ribosyltransferases that catalyzes polyadenosine diphosphate ribose (ADPR) formation in order to attract the DNA repair machinery to sites of DNA damage. The inhibition of PARP activity by olaparib can cause cell death, which is of clinical relevance in some tumor types. This demonstrates that quantification of PARP activity in the context of living cells is of great importance. In this work, we present the design, synthesis and biological evaluation of photo‐activatable affinity probes inspired by the olaparib molecule that are equipped with a diazirine for covalent attachment upon activation by UV light and a ligation handle for the addition of a reporter group of choice. SDS‐PAGE, western blotting and label‐free LC‐MS/MS quantification analysis show that the probes target the PARP‐1 protein and are selectively outcompeted by olaparib; this suggests that they bind in the same enzymatic pocket. Proteomics data are available via ProteomeXchange with identifier PXD018661. John Wiley and Sons Inc. 2020-05-13 2020-09-01 /pmc/articles/PMC7496120/ /pubmed/32282108 http://dx.doi.org/10.1002/cbic.202000042 Text en © 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Communications
Voorneveld, Jim
Florea, Bogdan I.
Bakkum, Thomas
Mendowicz, Rafal J.
van der Veer, Miriam S.
Gagestein, Berend
van Kasteren, Sander I.
van der Stelt, Mario
Overkleeft, Herman S.
Filippov, Dmitri V.
Olaparib‐Based Photoaffinity Probes for PARP‐1 Detection in Living Cells
title Olaparib‐Based Photoaffinity Probes for PARP‐1 Detection in Living Cells
title_full Olaparib‐Based Photoaffinity Probes for PARP‐1 Detection in Living Cells
title_fullStr Olaparib‐Based Photoaffinity Probes for PARP‐1 Detection in Living Cells
title_full_unstemmed Olaparib‐Based Photoaffinity Probes for PARP‐1 Detection in Living Cells
title_short Olaparib‐Based Photoaffinity Probes for PARP‐1 Detection in Living Cells
title_sort olaparib‐based photoaffinity probes for parp‐1 detection in living cells
topic Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7496120/
https://www.ncbi.nlm.nih.gov/pubmed/32282108
http://dx.doi.org/10.1002/cbic.202000042
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