Inhibition of hyaluronan hydrolysis catalysed by hyaluronidase at high substrate concentration and low ionic strength.
Hyaluronidase and high levels of hyaluronan are found together in tumours. It is highly likely that hyaluronidase activity controls the balance between high molecular mass hyaluronan and oligosaccharides, and thus plays an important role in cancer development. The hyaluronan hydrolysis catalysed by bovine testicular hyaluronidase was studied as a model. The kinetics was investigated at pH 5 and 37 degrees C using the colorimetric N-acetyl-d-glucosamine reducing end assay method. While the substrate dependence obtained in the presence of 0.15 mol L(-1) ionic strength exhibited a Michaelis-Menten behaviour, an atypical behaviour was observed under low ionic strength: for increasing hyaluronan concentrations, the initial reaction rate increased, reached a maximum and then decreased to a very low level, close to zero at high substrate concentrations. One of the various hypotheses examined to explain this atypical behaviour is the formation of non-specific complexes between hyaluronan and hyaluronidase based on electrostatic interactions. This hypothesis is the only one that can explain all the experimental results including the variation of the reaction medium turbidity as a function of time and the influence on the initial reaction rate of the hyaluronan concentration over hyaluronidase concentration. However, phenomena such as the high viscosity of highly concentrated hyaluronan solutions or the steric exclusion of hyaluronidase from hyaluronan solutions may contribute to the atypical behaviour. Finally, the biological implications of the non-linear and non-monotonous shape of the hyaluronan-hyaluronidase substrate dependence in the regulation of the hyaluronan chain molecular mass are discussed, in particular in the case of cancer development.
Astériou
Trias
T
Laboratoire Polymères, Biopolymères, Membranes, UMR 6522 Université de Rouen-CNRS, 76821 Mont-Saint-Aignan cedex, France.
Vincent
Jean-Claude
JC
Tranchepain
Frédéric
F
Deschrevel
Brigitte
B
D000818:Animals; D002417:Cattle; D006820:Hyaluronic Acid / Q000378:metabolism*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism*; D008297:Male; D009994:Osmolar Concentration; D013737:Testis / Q000201:enzymology
D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase