Affimer-mediated locking of p21-activated kinase 5 in an intermediate activation state results in kinase inhibition.
Cell Rep
; 42(10): 113184, 2023 10 31.
Article
in En
| MEDLINE
| ID: mdl-37776520
ABSTRACT
Kinases are important therapeutic targets, and their inhibitors are classified according to their mechanism of action, which range from blocking ATP binding to covalent inhibition. Here, a mechanism of inhibition is highlighted by capturing p21-activated kinase 5 (PAK5) in an intermediate state of activation using an Affimer reagent that binds in the P+1 pocket. PAK5 was identified from a non-hypothesis-driven high-content imaging RNAi screen in urothelial cancer cells. Silencing of PAK5 resulted in reduced cell number, G1/S arrest, and enlargement of cells, suggesting it to be important in urothelial cancer cell line survival and proliferation. Affimer reagents were isolated to identify mechanisms of inhibition. The Affimer PAK5-Af17 recapitulated the phenotype seen with siRNA. Co-crystallization revealed that PAK5-Af17 bound in the P+1 pocket of PAK5, locking the kinase into a partial activation state. This mechanism of inhibition indicates that another class of kinase inhibitors is possible.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
P21-Activated Kinases
/
Neoplasms
Limits:
Humans
Language:
En
Journal:
Cell Rep
Year:
2023
Document type:
Article