AtaGenix Laboratories
AtaGenix Laboratories
Release time: 2025-10-13 View volume: 327
Project Snapshot — A Cell Death & Disease study from Jiangxi Cancer Hospital revealed the mechanism by which ROCK2 promotes Pemigatinib resistance in cholangiocarcinoma (CCA) cells through UBA52-mediated DRP1 ubiquitination and ferroptosis regulation. AtaGenix provided four custom proteins (GST, GST-UBA52, His-ROCK2, His-DRP1) that enabled GST pull-down validation of the ROCK2/UBA52/DRP1 interaction axis — the core mechanistic evidence for this drug resistance pathway.
Based on: Cell Death & Disease, 2025 — DOI: 10.1038/s41419-025-07804-9
Cholangiocarcinoma (CCA) is a highly malignant tumor originating from the biliary epithelium, characterized by insidious onset and poor prognosis, with a five-year survival rate below 5%. In 2020, Pemigatinib became the first FDA-approved targeted therapy for CCA; however, clinical resistance frequently occurs, severely affecting therapeutic outcomes. Recent studies have identified Rho-associated coiled-coil containing protein kinase 2 (ROCK2) as a key regulator in chemotherapy resistance across various cancers.
Fig. 1. Mechanistic overview of ROCK2-mediated Pemigatinib resistance via the UBA52/DRP1 axis. Adapted from Cell Death Dis 2025.
Multi-Domain Protein Expression: UBA52 is a ubiquitin-related protein, while ROCK2 and DRP1 are multi-domain regulatory molecules. Their full-length or key domains are difficult to express in both prokaryotic and eukaryotic systems, often leading to misfolding, inclusion body formation, or partial degradation.
Indirect Interaction Validation: ROCK2 and DRP1 do not interact directly; rather, UBA52 mediates a competitive binding mechanism that regulates DRP1 ubiquitination. This poses significant challenges for in vitro reconstitution of the protein complex.
Post-Translational Modification: The enzymatic function of UBA52, an E3 ubiquitin ligase, depends on its proper conformation and activity. In vitro assays must ensure it can effectively mediate DRP1 ubiquitination.
Based on the client's requirements and provided sequences, AtaGenix delivered high-purity, low-endotoxin, correctly folded, and bioactive GST, GST-UBA52, His-ROCK2, and His-DRP1 proteins. These proteins played a pivotal role in Co-IP and Western blot analyses, helping elucidate that ROCK2 stabilizes DRP1 by competitively binding UBA52 — supporting the central role of the ROCK2/UBA52/DRP1 axis in CCA drug resistance mechanisms.
Fig. 2. GST pull-down and Western blot validation of ROCK2/UBA52/DRP1 protein interactions using AtaGenix custom proteins. Adapted from Cell Death Dis 2025.
4
Custom Proteins Delivered
GST + His
Tag Systems Used
Co-IP + WB
Validated Applications
CCA
Disease Model
Why This Matters
This study identifies the ROCK2/UBA52/DRP1 axis as a mechanistic driver of Pemigatinib resistance in CCA — providing both a resistance biomarker concept and a potential therapeutic intervention point. The AtaGenix custom proteins were essential for in vitro reconstitution of this multi-protein interaction: correctly folded, bioactive GST-UBA52 and His-ROCK2/His-DRP1 enabled the GST pull-down experiments that confirmed UBA52-mediated competitive binding as the core resistance mechanism.
Cell Death & Disease, 2025. DOI: 10.1038/s41419-025-07804-9
For research use only. Results may vary depending on target protein, expression system, and project scope. All proprietary client information is subject to NDA.
Need custom recombinant proteins for protein interaction validation, pull-down assays, or drug resistance mechanism studies? AtaGenix delivers high-purity, bioactive proteins with tag-specific purification and application-level QC.
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