A ubiquitination-mediated degradation system to target 14-3-3-binding phosphoproteins

Zhaokai Li; Xiaoqiang Huang; Mohan Li; Y Eugene Chen; Zhong Wang; Liu Liu

doi:10.1016/j.heliyon.2023.e16318

A ubiquitination-mediated degradation system to target 14-3-3-binding phosphoproteins

Heliyon. 2023 May 20;9(5):e16318. doi: 10.1016/j.heliyon.2023.e16318. eCollection 2023 May.

Authors

Zhaokai Li^{1

2}, Xiaoqiang Huang³, Mohan Li⁴, Y Eugene Chen^{1

3

5}, Zhong Wang¹, Liu Liu¹

Affiliations

¹ Department of Cardiac Surgery, Cardiovascular Center, University of Michigan, Ann Arbor, MI 48109, USA.
² Department of Emergency Medicine, China-Japan Friendship Hospital, Beijing 100029, China.
³ Center for Advanced Models for Translational Sciences and Therapeutics, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109, USA.
⁴ Department of Geriatrics, Chinese Academy of Medical Sciences, Peking Union Medical College Hospital, Beijing 100730, China.
⁵ Department of Internal Medicine, Cardiovascular Center, University of Michigan, Ann Arbor, MI 48109, USA.

Abstract

The phosphorylation of 14-3-3 binding motif is involved in many cellular processes. A strategy that enables targeted degradation of 14-3-3-binding phosphoproteins (14-3-3-BPPs) for studying their functions is highly desirable for basic research. Here, we report a phosphorylation-induced, ubiquitin-proteasome-system-mediated targeted protein degradation (TPD) strategy that allows specific degradation of 14-3-3-BPPs. Specifically, by ligating a modified von Hippel-Lindau E3-ligase with an engineered 14-3-3 bait, we generated a protein chimera referred to as Targeted Degradation of 14-3-3-binding PhosphoProtein (TDPP). TDPP can serve as a universal degrader for 14-3-3-BPPs based on the specific recognition of the phosphorylation in 14-3-3 binding motifs. TDPP shows high efficiency and specificity to a difopein-EGFP reporter, general and specific 14-3-3-BPPs. TDPP can also be applied for the validation of 14-3-3-BPPs. These results strongly support TDPP as a powerful tool for 14-3-3 related research.

Keywords: 14-3-3; Phosphorylation; Targeted protein degradation; Ubiquitination; von hippel-lindau.

Grants and funding

R01 HL163672/HL/NHLBI NIH HHS/United States