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).
| Original language | English |
|---|---|
| Article number | 4867 |
| Journal | Molecules |
| Volume | 27 |
| Issue number | 15 |
| DOIs | |
| State | Published - 1 Aug 2022 |
| Externally published | Yes |
Keywords
- antibody off-targets
- cell cycle
- posttranslational modifications
ASJC Scopus subject areas
- Drug Discovery
- Analytical Chemistry
- Chemistry (miscellaneous)
- Molecular Medicine
- Physical and Theoretical Chemistry
- Pharmaceutical Science
- Organic Chemistry