Identification and targeting of a novel drug pocket in the tumour angiogenesis regulator RhoJ
- 1School of Biochemistry and Biomedical Sciences, University of Bristol
Abstract
The Rho GTPase RhoJ is a key regulator of tumour angiogenesis and has been identified as a target for anti-angiogenic drug design. Its high sequence conservation with the related Rho GTPase Cdc42 complicates selective drug discovery. RhoJ has a unique N-terminal extension (Asn2-Lys19) that is absent in Cdc42 and other Rho GTPases. Here, we use molecular dynamics simulations to identify a novel allosteric pocket formed by the RhoJ N-terminus and show that this pocket contributes to interactions with key downstream signalling partners. Structure-based virtual screening of small molecules targeting this pocket identified the antifungal terconazole as a potential binder. Computational docking revealed that terconazole occupies the N-terminal pocket, sharing critical contacts with the interface between RhoJ and PAK3, a key interacting partner. To validate this, we developed an optimized bioluminescence resonance energy transfer (eBRET2)-based biosensor to monitor the interaction between RhoJ and PAK3 in live cells and saw that terconazole disrupted the RhoJ:PAK3 interaction in a dose-dependent manner. Together, our MD results show that the RhoJ N-terminal extension assumes structure on binding to PAK3 and contributes to that interaction, while our cellular biosensor data support targeting this pocket to modulate the RhoJ:PAK3 interaction. The ability of terconazole to bind to the N-terminal pocket and disrupt PAK3 binding provides proof of concept for the specific targeting of RhoJ in the development of novel anti-angiogenic drugs.
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BibTeX
@misc{chen2026identification,
title = {{Identification and targeting of a novel drug pocket in the tumour angiogenesis regulator RhoJ}},
author = {W. Chen and D. K. Shoemark and R. A. Corey and T. N. Adrienko and H. Mellor},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.14.758331},
url = {https://www.biorxiv.org/content/10.64898/2026.10.14.758331v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Identification and targeting of a novel drug pocket in the tumour angiogenesis regulator RhoJ
AU - W. Chen
AU - D. K. Shoemark
AU - R. A. Corey
AU - T. N. Adrienko
AU - H. Mellor
PY - 2026
DA - 2026/10/14
PB - bioRxiv
DO - 10.64898/2026.10.14.758331
UR - https://www.biorxiv.org/content/10.64898/2026.10.14.758331v1
AB - The Rho GTPase RhoJ is a key regulator of tumour angiogenesis and has been identified as a target for anti-angiogenic drug design. Its high sequence conservation with the related Rho GTPase Cdc42 complicates selective drug discovery. RhoJ has a unique N-terminal extension (Asn2-Lys19) that is absent in Cdc42 and other Rho GTPases. Here, we use molecular dynamics simulations to identify a novel allosteric pocket formed by the RhoJ N-terminus and show that this pocket contributes to interactions with key downstream signalling partners. Structure-based virtual screening of small molecules targeting this pocket identified the antifungal terconazole as a potential binder. Computational docking revealed that terconazole occupies the N-terminal pocket, sharing critical contacts with the interface between RhoJ and PAK3, a key interacting partner. To validate this, we developed an optimized bioluminescence resonance energy transfer (eBRET2)-based biosensor to monitor the interaction between RhoJ and PAK3 in live cells and saw that terconazole disrupted the RhoJ:PAK3 interaction in a dose-dependent manner. Together, our MD results show that the RhoJ N-terminal extension assumes structure on binding to PAK3 and contributes to that interaction, while our cellular biosensor data support targeting this pocket to modulate the RhoJ:PAK3 interaction. The ability of terconazole to bind to the N-terminal pocket and disrupt PAK3 binding provides proof of concept for the specific targeting of RhoJ in the development of novel anti-angiogenic drugs.
ER -