Detecting protein ADP-ribosylation using a clickable aminooxy probe

Rory K. Morgan, Michael Cohen

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

ADP-ribosylation, a posttranslational modification catalyzed by a family of enzymes known as poly(ADP-ribose) polymerases (PARPs, 17 in humans), regulates diverse cellular processes. To aid in understanding the functions of ADP-ribosylation in cells, we developed a clickable aminooxy probe, AO-alkyne, which detects ADP-ribosylation of acidic amino acids. AO-alkyne can be used to detect auto-ADP-ribosylation of PARP10 in cells following Cu-catalyzed click conjugation to an azide reporter. This method can be extended to other PARP family members that catalyze ADP-ribosylation on acidic amino acids, providing a convenient and direct readout of PARP activity in cells.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages71-77
Number of pages7
Volume1608
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1608
ISSN (Print)1064-3745

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Adenosine Diphosphate
Acidic Amino Acids
Alkynes
Proteins
Azides
Poly(ADP-ribose) Polymerases
Post Translational Protein Processing
Enzymes

Keywords

  • ADP-ribosylation
  • Bioconjugation
  • Click chemistry
  • Oxime ligation
  • PARPs

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Morgan, R. K., & Cohen, M. (2017). Detecting protein ADP-ribosylation using a clickable aminooxy probe. In Methods in Molecular Biology (Vol. 1608, pp. 71-77). (Methods in Molecular Biology; Vol. 1608). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6993-7_6

Detecting protein ADP-ribosylation using a clickable aminooxy probe. / Morgan, Rory K.; Cohen, Michael.

Methods in Molecular Biology. Vol. 1608 Humana Press Inc., 2017. p. 71-77 (Methods in Molecular Biology; Vol. 1608).

Research output: Chapter in Book/Report/Conference proceedingChapter

Morgan, RK & Cohen, M 2017, Detecting protein ADP-ribosylation using a clickable aminooxy probe. in Methods in Molecular Biology. vol. 1608, Methods in Molecular Biology, vol. 1608, Humana Press Inc., pp. 71-77. https://doi.org/10.1007/978-1-4939-6993-7_6
Morgan RK, Cohen M. Detecting protein ADP-ribosylation using a clickable aminooxy probe. In Methods in Molecular Biology. Vol. 1608. Humana Press Inc. 2017. p. 71-77. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-6993-7_6
Morgan, Rory K. ; Cohen, Michael. / Detecting protein ADP-ribosylation using a clickable aminooxy probe. Methods in Molecular Biology. Vol. 1608 Humana Press Inc., 2017. pp. 71-77 (Methods in Molecular Biology).
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