RRC ID 68871
Author Tsumagari K, Chang CH, Ishihama Y.
Title A protocol for analyzing the protein terminome of human cancer cell line culture supernatants.
Journal STAR Protoc
Abstract Characterization of protein termini is essential for understanding how the proteome is generated through biological processes such as post-translational proteolytic events. Here, we introduce a practical protocol for terminomics to achieve simple and sensitive N- and C-terminal peptide enrichment. We apply it to the terminome analysis of culture supernatants of a human cancer cell line for the purpose of identifying ectodomain shedding substrate cleavage sites with 10 μg protein per sample. For complete details on the use and execution of this protocol, please refer to Tsumagari et al. (2021).
Volume 2(3)
Pages 100682
Published 2021-9-17
DOI 10.1016/j.xpro.2021.100682
PII S2666-1667(21)00389-0
PMID 34377995
PMC PMC8327658
MeSH Cell Line, Tumor / metabolism Humans Isotope Labeling / methods* Peptide Hydrolases / metabolism Peptides / chemistry Protein Domains / physiology* Protein Processing, Post-Translational / physiology Proteolysis / drug effects Proteome / analysis Proteomics / methods* Tandem Mass Spectrometry / methods
Resource
Human and Animal Cells A431(RCB0202)