学术文献库

${\bf Background}$ 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.