In this section
@article{Kalesh:2019:10.1038/s41598-019-43154-1,
author = {Kalesh, K and Lukauskas, S and Borg, AJ and Snijders, AP and Ayyappan, V and Leung, AKL and Haskard, DO and DiMaggio, PA},
doi = {10.1038/s41598-019-43154-1},
journal = {Scientific Reports},
pages = {1--12},
title = {An integrated chemical proteomics approach for quantitative profiling of intracellular ADP-Ribosylation},
url = {http://dx.doi.org/10.1038/s41598-019-43154-1},
volume = {9},
year = {2019}
}
TY - JOUR
AB - ADP-ribosylation is integral to a diverse range of cellular processes such as DNA repair, chromatin regulation and RNA processing. However, proteome-wide investigation of its cellular functions has been limited due to numerous technical challenges including the complexity of the poly(ADP-ribose) (PAR) chains, low abundance of the modification and lack of sensitive enrichment methods. We herein show that an adenosine analogue with a terminal alkyne functionality at position 2 of the adenine (2-alkyne adenosine or 2YnAd) is suitable for selective enrichment, fluorescence detection and mass spectrometry proteomics analysis of the candidate ADP-ribosylome in mammalian cells. Although similar labelling profiles were observed via fluorescence imaging for 2YnAd and 6YnAd, a previously reported clickable NAD+ precursor, quantitative mass spectrometry analysis of the two probes in MDA-MB-231 breast cancer cells revealed a significant increase in protein coverage of the 2YnAd probe. To facilitate global enrichment of ADP-ribosylated proteins, we developed a dual metabolic labelling approach that involves simultaneous treatment of live cells with both 2YnAd and 6YnAd. By combining this dual metabolic labelling strategy with highly sensitive tandem mass tag (TMT) isobaric mass spectrometry and hierarchical Bayesian analysis, we have quantified the responses of thousands of endogenous proteins to clinical PARP inhibitors Olaparib and Rucaparib.
AU - Kalesh,K
AU - Lukauskas,S
AU - Borg,AJ
AU - Snijders,AP
AU - Ayyappan,V
AU - Leung,AKL
AU - Haskard,DO
AU - DiMaggio,PA
DO - 10.1038/s41598-019-43154-1
EP - 12
PY - 2019///
SN - 2045-2322
SP - 1
TI - An integrated chemical proteomics approach for quantitative profiling of intracellular ADP-Ribosylation
T2 - Scientific Reports
UR - http://dx.doi.org/10.1038/s41598-019-43154-1
UR - https://www.ncbi.nlm.nih.gov/pubmed/31040352
UR - https://www.nature.com/articles/s41598-019-43154-1
VL - 9
ER -