Protocol for the quantitative identification of endogenously ISGylated proteins from mammalian cell lines Journal Article

   


Authors: Wardlaw, C. P.; Miele, M. M.; Li, Z.; Hendrickson, R. C.; Petrini, J. H. J.
Article Title: Protocol for the quantitative identification of endogenously ISGylated proteins from mammalian cell lines
Abstract: Ubiquitin-like protein ISG15 plays an important role in an array of cellular functions via its covalent attachment to target proteins (ISGylation). Here, we present a protocol for the identification of ISGylated proteins that avoids the caveats associated with ISG15 overexpression and minimizes the likelihood of false positives. We describe steps for the tagging of endogenous ISG15, followed by genotyping and clone selection. We then detail steps for ISGylation induction, the isolation of ISGylated proteins, and their identification via quantitative mass spectrometry. For complete details on the use and execution of this protocol, please refer to Wardlaw and Petrini.1 © 2024 The Author(s)
Keywords: unclassified drug; nonhuman; ubiquitin; polymerase chain reaction; mass spectrometry; dna damage; protein; proteomics; western blotting; molecular biology; dna extraction; fluorescence activated cell sorting; mammal cell; ribonucleotide; cell biology; genotyping; dna template; sanger sequencing; dna sequencing; article; ultra performance liquid chromatography; crispr; clustered regularly interspaced short palindromic repeat; electronic spreadsheet; isgylated proteins
Journal Title: STAR Protocols
Volume: 5
Issue: 1
ISSN: 2666-1667
Publisher: Cell Press  
Date Published: 2024-03-15
Start Page: 102843
Language: English
DOI: 10.1016/j.xpro.2024.102843
PROVIDER: scopus
PMCID: PMC10844860
PUBMED: 38294909
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85183526527&doi=10.1016%2fj.xpro.2024.102843&partnerID=40&md5=b9323c6c9d31a0048aad05f8fb479bcb
Notes: The MSK Cancer Center Support Grant (P30 CA008748) is acknowledged in the PDF -- Corresponding author is MSK author: John H.J. Petrini -- Source: Scopus
Altmetric
What is Altmetric?
Citation Impact
What is Dimensions Citation Badge?
BMJ Impact Analytics
MSK Authors
  1. John Petrini
    94 Petrini
  2. Ronald Charles Hendrickson
    86 Hendrickson
  3. Matthew M Miele
    18 Miele
  4. Zhuoning Li
    17 Li
  5. Christopher Wardlaw
    3 Wardlaw
Related MSK Work