pSNAP: Proteome-wide analysis of elongating nascent polypeptide chains

• DOI: 10.1016/j.isci.2022.104516
• Publication Date: 2022-06-03T01:47:18Z
• Authors: Junki Uchiyama; Rohini Roy; Dan Ohtan Wang; Kazuya Morikawa; Yuka Kawahara; Mio Iwasaki; Chiaki Yoshino; Yuichiro Mishima; Yasushi Ishihama; Koshi Imami
• Keywords: Proteome; Ribosome profiling; Puromycin

Cellular global translation is often measured using ribosome profiling or quantitative mass spectrometry, but these methods do not provide direct information at the level of elongating nascent polypeptide chains (NPCs) and associated co-translational events. Here, we describe pSNAP, a method for proteome-wide NPCs by affinity enrichment puromycin- stable isotope-labeled polypeptides. pSNAP does require purification and/or chemical labeling, captures bona fide that characteristically exhibit protein N-terminus-biased positions. We applied to evaluate effect silmitasertib, potential molecular therapy cancer, revealed acute translational repression through casein kinase II mTOR pathways. also characterized modifications on demonstrated combination different types modifications, such as acetylation phosphorylation in N-terminal region histone H1.5, can modulate interactions with ribosome-associated factors. Thus, provides framework dissecting regulations scale.