Computational investigation of tubulin mutations effect on colchicine binding site

Colchicine derivatives are currently under scrutiny as potential candidates for future cancer therapeutic applications, leveraging the well-established antimitotic properties of colchicine. In this study, a comprehensive computational model of the colchicine binding site has been constructed within tubulin heterodimers. To achieve this, AlphaFold2 has been employed to generate three-dimensional structures of human beta tubulin in its wildtype and mutated form. A human alpha tubulin structure from available data bank online has been used to create the heterodimer, subsequently modifying the geometry of the heterodimer incorporating colchicine. This homology modeling approach was guided by crystallographic data from colchicine-docked microtubules in bovine cells. This computational model was developed for various forms of the heterodimer, with a particular focus on different isotypes of beta tubulin that are known to be overexpressed in cancer tissues. To validate the model's accuracy and reliability, a rigorous comparison with experimental data derived from animal studies has been conducted. Furthermore, the computational models were harnessed to perform docking simulations with colchicine, focusing on the variations in binding affinity induced by specific mutations within the colchicine binding site. This comprehensive analysis provides critical insights into the impact of distinct beta tubulin mutations on the colchicine binding site and offers valuable information essential for the design and development of highly targeted colchicine-based drugs tailored to tubulins overexpressed in cancer tissues. These findings represent a promising step towards the creation of more effective and specific cancer therapeutics.