| Abstract: | Competition experiments were carried out on the hydrolysis of different substrates by beta-hexosaminidase A isolated from human liver. The results show that ganglioside GM2 in the presence of the GM2 activator protein and a new synthetic substrate, 4-methylumbelliferyl-beta-N-acetylglucosaminide 6-sulfate, are hydrolyzed at the same active site on the alpha subunit of beta-hexosaminidase A, whereas 4-methylumbelliferyl-beta-N-acetylglucosaminide is degraded predominantly by a different active site on the beta-subunit. This finding provides for the first time a possible molecular basis for the observation that, in variant B1 of the GM2 gangliosidoses, beta-hexosaminidase A has lost its activity toward GM2 ganglioside and the sulfated artificial substrate while being still able to hydrolyze the unsulfated artificial substrate at a normal rate. Furthermore, the finding that the GM2 activator protein inhibits the degradation of the sulfated substrate by beta-hexosaminidases A and S indicates that the alpha subunit common to both isoenzymes might provide a binding site for the activator protein. |
