Methodological insights into the assembly and validation of USP22 complexes via the PQLink vector suite

Zhengyang Zhao; Feng Wang

doi:10.1117/12.3044075

27 August 2024 Methodological insights into the assembly and validation of USP22 complexes via the PQLink vector suite

Zhengyang Zhao, Feng Wang

Proceedings Volume 13252, Fourth International Conference on Biomedicine and Bioinformatics Engineering (ICBBE 2024); 132520Y (2024) https://doi.org/10.1117/12.3044075
Event: 2024 Fourth International Conference on Biomedicine and Bioinformatics Engineering (ICBBE 2024), 2024, Kaifeng, China

Abstract

USP22, as a deubiquitinating enzyme, is highly expressed in various types of cancer and promotes the progression of the cell cycle and cancer development by interacting with cell cycle regulatory factors. This study, for the first time, successfully expressed a highly active USP22 complex in vitro using a method based on the PQLink series of vectors. It was found that USP22 must form a complex with the auxiliary proteins ATXN7, ATXN7L3, and ENY2 to possess deubiquitinating enzyme activity. Through optimizing gene sequences, the construction of protein expression vectors, expression-test, overexpression and purification, we obtained a highly active USP22 complex and verified its deubiquitinating enzyme activity using Ub-AMC hydrolysis assay. In addition, the experimental results showed that the enzyme activity of the USP22 complex would be lost if any of the auxiliary proteins were missing. This study not only provides important information for understanding the mechanism of USP22 in cancer but also lays the foundation for the development of small molecule inhibitors targeting USP22, which is expected to become a new strategy for cancer treatment.

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

Citation Download Citation

Zhengyang Zhao and Feng Wang "Methodological insights into the assembly and validation of USP22 complexes via the PQLink vector suite", Proc. SPIE 13252, Fourth International Conference on Biomedicine and Bioinformatics Engineering (ICBBE 2024), 132520Y (27 August 2024); https://doi.org/10.1117/12.3044075

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available