Kinetic investigation of the action of hyaluronidase on hyaluronan using the Morgan-Elson and neocuproine assays.
In this paper we describe kinetic investigations of the action of testicular hyaluronidase on hyaluronan. We have compared the use of two spectrophotometric assays, the first based on the Morgan-Elson reaction and the second on the neocuproine reaction. With the neocuproine reaction Km was found to be 0.46 mg/ml and Vmax to be 126 nmol l-1 s-1. Because of a low sensitivity and the production of interfering precipitates, the Morgan-Elson assay cannot be used for kinetic investigation of the enzyme. Furthermore this assay is prone to interference from compounds such as disodium cromoglycate, (+)-catechine, penicillamine, CaCl2 and acetate buffer.
Vercruysse
K P
KP
Laboratory of General Biochemistry, University of Ghent, Belgium.
Lauwers
A R
AR
Demeester
J M
JM
D000117:Acetylglucosamine / Q000378:metabolism; D002240:Carbohydrate Sequence; D002614:Chelating Agents / Q000737:chemistry; D006820:Hyaluronic Acid / Q000378:metabolism*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D010618:Phenanthrolines / Q000737:chemistry*; D015203:Reproducibility of Results
D002614:Chelating Agents; D010618:Phenanthrolines; C002701:neocuproine; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase; D000117:Acetylglucosamine