| Products/Services Used | Details | Operation |
|---|---|---|
| Custom Vector Construction | Human HPF1 (pET21a), human RPS3A-WT (pGEX6P-3), and human RPS3A-Y155F (pGEX6P-3) bacterial expression vectors were constructed by GenScript (Piscataway, NJ, USA). Plasmids were transformed into BL21 Escherichia coli, and protein expression was induced by adding 1 mM IPTG at OD600 0.4–0.6 for 3 h at 30°C. | Get A Quote |
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 ADPr-acceptor amino acid. MS analyses of ADP-ribosylated proteins confirmed tyrosine as an ADPr-acceptor amino acid in RPS3A (Y155) and HPF1 (Y238) and demonstrated that -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 ... More
