Production of phosphorylated beta-catenin by protein semi-synthesis
Embargo Date
2028-02-10
OA Version
Citation
Abstract
Beta-catenin, a transcription factor involved in the wingless-related integration site (WNT) signaling pathway, has been linked to many different cancers such clear cell renal carcinoma and colorectal cancer. Dysregulation of Beta-catenin/WNT signaling pathway leads to uncontrolled proliferation, differentiation, and inhibition of apoptosis. Given that beta-catenin serves a central role in WNT signaling pathway, it is subject to tight regulation including by post-translational modifications. Phosphorylation of beta-catenin in its N- terminal segment at Ser33 and Ser37 by glycogen synthase kinase 3 (GSK3) regulates its activity, localization, and stability by recruiting the beta-transducin repeat-containing protein (beta-TrCP) E3 ubiquitin ligase complex to ubiquitinate beta-catenin leading to its proteasomal destruction. Moreover, there are other phosphorylation sites in this segment and throughout beta-catenin that have not been fully characterized. Herein, we used expressed protein ligation to generate di-phosphorylated beta-catenin at Ser33 and Ser37 along with fusing a biotin ligase known as biotin-based identification interacting protein 2 (BioID2) to its C-terminus as a proof of concept to generate a new protein regent to discover and assess other protein binding partners.
Description
2025