Proteomics

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Proteome-wide analysis of elongating nascent polypeptide chains – LPS stimulation to RAW264.7 macrophages

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. The inputs (without IP) were analyzed in triplicates.

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Yasushi Ishihama 

PROVIDER: PXD031475 | JPOST Repository | Tue Feb 07 00:00:00 GMT 2023

REPOSITORIES: jPOST

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211014KI_225_MM_LPS60min_H120min_1.raw Raw
211014KI_225_MM_LPS60min_H120min_2.raw Raw
211014KI_225_MM_LPS60min_H120min_3.raw Raw
211014KI_225_MM_LPS60min_H30min_1.raw Raw
211014KI_225_MM_LPS60min_H30min_2.raw Raw
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Publications

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]

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