Proteomics

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Proteome-wide analysis of elongating nascent polypeptide chains - mouse primary neuron

ABSTRACT: 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. We combine quantitative proteomics, pulse labeling with puromycin, and stable isotope-labeled amino acids to analyze thousands of NPCs. Our approach enables global analyses of translational responses and co-translational modifications, providing a framework for dissecting co-translational regulations proteome-wide. NPCs from mouse primary neurons were analyzed in triplicates.

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Yasushi Ishihama 

PROVIDER: PXD024104 | JPOST Repository | Tue Feb 07 00:00:00 GMT 2023

REPOSITORIES: jPOST

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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 profiling of NPCs by affinity enrichment of puromycin- and stable isotope-labeled polypeptides. pSNAP does not require ribosome purification and/or chemical labeling, and captures <i>bona f  ...[more]