Deciphering Mechanism of Chondroitinase ABC Copolymer Stabilization Via Molecular Dynamics

Abstract

Chondroitinase ABC is an enzyme that has shown therapeutic potential in treating spinal cord injuries through the breakdown of glial scarring, which inhibits axonal regrowth. However, it is unstable at human body temperature, and has been difficult to stabilize, which makes it challenging to implement as a therapeutic. Thus, a collaboration between the Webb and Gormley labs identified and tested the ability of copolymers to serve as stabilizing agents. Many of these copolymers successfully boosted the retained enzyme activity (REA) of chABC over 24 hours at 310K, or human body temperature. However, the experiments involved in this study did not reveal the mechanism by which these copolymers endowed thermodynamic stability. If this mechanism of stabilization were better understood, it would contribute to the understanding of chABC’s thermal instabilities and forward the goal of implementing chABC as a therapeutic. Molecular dynamics simulations were utilized to computationally interpret the mechanism of copolymer stabilization of chABC. These simulations revealed that there are facets of chABC’s wet lab behavior that can be understood and interpreted computationally, as well as contribute to the understanding of copolymer chABC stabilization. Investigating the interactions between chABC and high REA copolymers revealed that high levels of intermolecular contact can lead to a maintained protein structure, while low REA copolymers can exhibit distinctly destabilizing behavior not exhibited in a laboratory setting. These impacts may be related to the ability of copolymers to block the collapse of chABC from a crescent into a torus. Successful blocking of destabilizing intra-protein interactions, along with high contact bracing, seems to result in the most successful stabilization of chABC. Conversely, blocking the destabilizing intra-protein interactions without any additional support results in the most significant destabilization of chABC.