Kinetic theory of Hyaluronan cleavage by Bovine Testicular Hyaluronidase in Standard and Crowded Environments

TL;DR

This study develops a detailed kinetic model for hyaluronan degradation by hyaluronidase, revealing key pathways and effects of crowding, providing new insights into enzymatic HA breakdown in physiological conditions.

Contribution

Introduces a comprehensive kinetic model for HA degradation, identifying dominant pathways and the impact of crowding on enzyme kinetics.

Findings

All relevant fragmentation and transglycosylation pathways identified

Recombination of two dimers into a tetramer is dominant

Crowding slows kinetics but does not change mechanisms

Abstract

${\bf Background}$ Details of the kinetic pathways governing enzymatic cleavage of hyaluronic acid (HA) by hyaluronidase are still widely uncharted. Capillary electrophoresis-based assays were used for accurate quantification of enzymatic products. A crowding agent was also employed to mimic excluded-volume constraints typical of in-vivo conditions. $ {\bf Scope}$ Introduce a comprehensive kinetic model describing the late-stage degradation of HA by hyaluronidase and identify the relevant kinetic pathways and the associated rates. ${\bf Major Conclusions}$ All relevant fragmentation and transglycosylation pathways and rates were identified. Two dimers forming a tetramer is the dominant recombination pathway. Macromolecular and self-crowding slow down the kinetics but do not alter the underlying mechanisms. ${\bf General Significance}$ Our results bring a novel and comprehensive quantitative insight into enzymatic HA degradation. Rationalizing the effect of crowding brings the intricate conditions of in-vivo settings a little closer, and also stands as a powerful tool to pinpoint relevant kinetic pathways in complex systems.