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Challenges 2018, 9(1), 23; https://doi.org/10.3390/challe9010023

In Vivo Level of Poly(ADP-ribose)

1
Nagahama Institute of Bio-Science and Technology, Nagahama, Shiga 526-0829, Japan
2
Department of Applied Life Sciences, College of Nagoya Women’s University, Nagoya-shi, Aichi 467-8610, Japan
3
Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-0046, Japan
4
Center for Chronic Viral Diseases, Kagoshima University, Sakuragaoka 8-35-1, Kagoshima 890-8544, Japan
*
Author to whom correspondence should be addressed.
Received: 10 April 2018 / Revised: 18 May 2018 / Accepted: 18 May 2018 / Published: 22 May 2018
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Abstract

PolyADP-ribosylation is a post-translational modification that plays key roles in cellular physiological functions and DNA damage responses. PolyADP-ribosylation is finely and dynamically regulated by various enzymes and factors involved in the synthesis and degradation of poly(ADP-ribose) (PAR). To better understand the function of polyADP-ribosylation, it is necessary to quantify and monitor the change of the in vivo level of PAR, the product of polyADP-ribosylation, which is rapidly turning over and kept in quite low level in cells or in organs. Recent developments of potent inhibitors of polyADP-ribosylation is expected to kill BRCA1/2-mutated breast cancer cells and ovarian cancer cells (synthetic lethality). To know the efficacy of these inhibitors in vivo, it is necessary to develop highly sensitive and reproducible methods to know PAR levels within cells or organs. However there have been several difficulties in measuring the physiologically low level of PAR without artefacts. Our experiments recently clarified that the method of sample preparation is very important in addition to the sensitivity and specificity. From reviewing the literature, including ours, we would like to emphasize the importance of the procedures of sample preparation for the assay, in addition to the sensitivity by comparing the reported PAR levels in vivo. View Full-Text
Keywords: polyADP-ribosylation; poly(ADP-ribose); PARP; PARG; PARP inhibitor; radioimmunoassay; ELISA; LC-MS/MS polyADP-ribosylation; poly(ADP-ribose); PARP; PARG; PARP inhibitor; radioimmunoassay; ELISA; LC-MS/MS
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Miwa, M.; Ida, C.; Yamashita, S.; Kouyama, K.; Kuroda, Y.; Eguchi, T.; Ohta, N.; Sato, T.; Tsuda, M.; Tanaka, M. In Vivo Level of Poly(ADP-ribose). Challenges 2018, 9, 23.

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