Antibody for Serine 65 Phosphorylated Ubiquitin Identifies PLK1-Mediated Phosphorylation of Mitotic Proteins and APC1.
Antibodies
/ genetics
Apc1 Subunit, Anaphase-Promoting Complex-Cyclosome
/ genetics
Cell Cycle Proteins
/ genetics
HeLa Cells
Humans
Mitosis
/ genetics
Phosphorylation
Protein Binding
/ genetics
Protein Processing, Post-Translational
/ genetics
Protein Serine-Threonine Kinases
/ genetics
Proto-Oncogene Proteins
/ genetics
Serine
/ genetics
Ubiquitin
/ genetics
Ubiquitination
Polo-Like Kinase 1
antibody off-targets
cell cycle
posttranslational modifications
Journal
Molecules (Basel, Switzerland)
ISSN: 1420-3049
Titre abrégé: Molecules
Pays: Switzerland
ID NLM: 100964009
Informations de publication
Date de publication:
29 Jul 2022
29 Jul 2022
Historique:
received:
23
06
2022
revised:
22
07
2022
accepted:
25
07
2022
entrez:
12
8
2022
pubmed:
13
8
2022
medline:
16
8
2022
Statut:
epublish
Résumé
Deciphering the protein posttranslational modification (PTM) code is one of the greatest biochemical challenges of our time. Phosphorylation and ubiquitylation are key PTMs that dictate protein function, recognition, sub-cellular localization, stability, turnover and fate. Hence, failures in their regulation leads to various disease. Chemical protein synthesis allows preparation of ubiquitinated and phosphorylated proteins to study their biochemical properties in great detail. However, monitoring these modifications in intact cells or in cell extracts mostly depends on antibodies, which often have off-target binding. Here, we report that the most widely used antibody for ubiquitin (Ub) phosphorylated at serine 65 (pUb) has significant off-targets that appear during mitosis. These off-targets are connected to polo-like kinase 1 (PLK1) mediated phosphorylation of cell cycle-related proteins and the anaphase promoting complex subunit 1 (APC1).
Identifiants
pubmed: 35956818
pii: molecules27154867
doi: 10.3390/molecules27154867
pmc: PMC9369648
pii:
doi:
Substances chimiques
ANAPC1 protein, human
0
Antibodies
0
Apc1 Subunit, Anaphase-Promoting Complex-Cyclosome
0
Cell Cycle Proteins
0
Proto-Oncogene Proteins
0
Ubiquitin
0
Serine
452VLY9402
Protein Serine-Threonine Kinases
EC 2.7.11.1
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : European Research Council
ID : 831783
Pays : International
Organisme : Israel Science Foundation
ID : 755/19
Organisme : Ministry of Science and Culture of Lower Saxony for research on mitochondria
ID : 76251-12-9/19 (ZN3457)
Organisme : ProteoCure
ID : CA20113
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