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Bottom-up and Shotgun Proteomics to Identify a Comprehensive Cochlear Proteome
Published on: March 7, 2014
Comprehensive ADP-ribosylome analysis identifies tyrosine as an ADP-ribose acceptor site
Deena M Leslie Pedrioli1, Mario Leutert1,2, Vera Bilan1
1Department of Molecular Mechanisms of Disease, University of Zurich, Zurich, Switzerland.
Abstract:
Despite recent mass spectrometry (MS)-based breakthroughs, comprehensive ADP-ribose (ADPr)-acceptor amino acid identification and ADPr-site localization remain challenging. Here, we report the establishment of an unbiased, multistep ADP-ribosylome data analysis workflow that led to the identification of tyrosine as a novel ARTD1/PARP1-dependent in vivo ADPr-acceptor amino acid. MS analyses of in vitro ADP-ribosylated proteins confirmed tyrosine as an ADPr-acceptor amino acid in RPS3A (Y155) and HPF1 (Y238) and demonstrated that trans-modification of RPS3A is dependent on HPF1. We provide an ADPr-site Localization Spectra Database (ADPr-LSD), which contains 288 high-quality ADPr-modified peptide spectra, to serve as ADPr spectral references for correct ADPr-site localizations.
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