Glucose 6-phosphate and mannose 6-phosphate are equally and more actively hydrolyzed by glucose 6-phosphatase during hysteretic transition within intact microsomal membrane than after detergent treatment.
We have studied the rapid kinetics of glucose-6-phosphatase (Glc6Pase) toward glucose 6-phosphate (Glc6P) and mannose 6-phosphate (Man6P) in intact and detergent-treated microsomes, using a radiometric assay based on the use of [U(-)14C]hexose 6-phosphates. We show that a hysteretic transition of Glc6Pase from a rapid hydrolytic form to slower kinetic form within the intact membrane occurs for both substrates with the same relaxation time. During the hysteretic transition, preceding the steady-state rate of hydrolysis, Glc6Pase is able to hydrolyze both Glc6P and Man6P at very similar rates. Only Glc6P is significantly hydrolyzed at steady state. Moreover, the initial rates of hydrolysis of both Glc6P and Man6P in intact microsomes are higher than the respective rates of hydrolysis after detergent treatment of the membrane at high substrate concentrations (10 and 20 microM), while these rates are not different at lower substrate concentrations. These data show that the marked specificity of Glc6Pase at steady state in the membrane is acquired owing to a hysteretic transition induced by the hydrolytic phenomenon, independently of the nature of the prior phosphate donor. The role of the membrane in this phenomenon is crucial, since the transition does not occur in its absence.
Ajzannay
A
A
Institut National de la Santé et de la Recherche Médicale, Unité 449, Lyon, France.
Mithieux
G
G
D000818:Animals; D003902:Detergents; D005952:Glucose-6-Phosphatase / Q000378:metabolism*; D019298:Glucose-6-Phosphate; D005958:Glucosephosphates / Q000378:metabolism*; D006868:Hydrolysis; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D008360:Mannosephosphates / Q000378:metabolism*; D008862:Microsomes, Liver / Q000378:metabolism*; D051381:Rats; D013379:Substrate Specificity
D003902:Detergents; D005958:Glucosephosphates; D008360:Mannosephosphates; C027693:mannose-6-phosphate; D019298:Glucose-6-Phosphate; D005952:Glucose-6-Phosphatase