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Antibody for Serine 65 Phosphorylated Ubiquitin Identifies PLK1-Mediated Phosphorylation of Mitotic Proteins and APC1

Authors :
Guy Mann
Prasad Sulkshane
Pradeep Sadhu
Tamar Ziv
Michael H. Glickman
Ashraf Brik
Source :
Molecules, Vol 27, Iss 15, p 4867 (2022)
Publication Year :
2022
Publisher :
MDPI AG, 2022.

Abstract

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).

Details

Language :
English
ISSN :
14203049
Volume :
27
Issue :
15
Database :
Directory of Open Access Journals
Journal :
Molecules
Publication Type :
Academic Journal
Accession number :
edsdoj.f5b12faaa7ff40d189963dd5bf099dbf
Document Type :
article
Full Text :
https://doi.org/10.3390/molecules27154867