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Detecting Protein ADP-Ribosylation Using a Clickable Aminooxy Probe
Rory K Morgan1, Michael S Cohen2
1Program in Chemical Biology, Department of Physiology and Pharmacology, Oregon Health and Science University, 3181 SW Sam Jackson Park Rd, #L334, Biomedical Research Building, Rm 621, Portland, OR, 97239-3098, USA.
Methods in Molecular Biology (Clifton, N.J.)
|July 12, 2017
Summary
Researchers developed a new clickable probe, AO-alkyne, to detect poly(ADP-ribose) polymerase (PARP) activity by visualizing ADP-ribosylation of acidic amino acids in cells.
Area of Science:
- Biochemistry
- Molecular Biology
- Cellular Biology
Background:
- ADP-ribosylation is a crucial posttranslational modification regulating cellular functions.
- Poly(ADP-ribose) polymerases (PARPs) catalyze ADP-ribosylation, with 17 identified in humans.
- Understanding PARP activity in cells is essential for deciphering cellular processes.
Purpose of the Study:
- To develop a novel chemical tool for detecting ADP-ribosylation.
- To enable visualization of PARP activity through a specific modification.
- To facilitate research into the diverse roles of ADP-ribosylation.
Main Methods:
- Development of a clickable aminooxy probe, AO-alkyne.
- Detection of ADP-ribosylation on acidic amino acids.
- Utilizing copper-catalyzed click chemistry for reporter conjugation.
Main Results:
- AO-alkyne successfully detected auto-ADP-ribosylation of PARP10 in cellular contexts.
- The method demonstrated effective detection following click conjugation to an azide reporter.
- The probe's utility was validated for PARP10 and is extendable to other PARP family members.
Conclusions:
- AO-alkyne provides a convenient and direct method for assessing PARP activity in cells.
- This probe advances the study of ADP-ribosylation and its regulatory roles.
- The developed technology offers a versatile tool for biochemical and cellular research.
