Thermal stability of microsomal glucose-6-phosphatase.
The thermal stability of glucose-6-phosphatase in rat liver microsomes was examined in untreated and cholate-treated microsomes. Activity of the enzyme was measured with both glucose-6-P and mannose-6-P as substrates. Heat treatment did not cause glucose-6-phosphatase activity to decline to zero with a single rate constant in untreated microsomes. Instead, heat treatment produced an enzyme with a small residual activity that was stable. The residual level of activity was not stimulated by addition of detergent. In untreated microsomes the energies of activation for the processes of decay were different for glucose-6-phosphatase and mannose-6-phosphatase activities, suggesting that the rate-limiting steps for the hydrolysis of these compounds were different. Treatment of microsomes with detergent increased the rate constants for the thermal decay of glucose-6-phosphatase by about 150 times, and, in contrast to untreated microsomes, glucose-6-phosphatase and mannose-6-phosphatase decayed to zero with a single rate constant in cholate-treated microsomes. Also, rate constants for thermal inactivation of glucose-6-phosphatase and mannose-6-phosphatase were the same in cholate-treated microsomes. Removal of cholate increased the stability of glucose-6-phosphatase but did not regenerate the form of the enzyme present in untreated microsomes. The data for the stability of glucose-6-phosphatase under different conditions provide evidence that the enzyme can exist in at least five different stable states that are enzymatically active.
Zakim
D
D
Division of Digestive Diseases, Cornell University Medical College, New York, New York 10021.
Dannenberg
A
A
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D019826:Cholic Acid; D002793:Cholic Acids / Q000494:pharmacology; D004355:Drug Stability; D005952:Glucose-6-Phosphatase / Q000378:metabolism*; D019298:Glucose-6-Phosphate; D005958:Glucosephosphates / Q000378:metabolism; D006358:Hot Temperature; D006868:Hydrolysis; D007700:Kinetics; D008297:Male; D008360:Mannosephosphates / Q000378:metabolism; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D013816:Thermodynamics
D002352:Carrier Proteins; D002793:Cholic Acids; D005958:Glucosephosphates; D008360:Mannosephosphates; C027693:mannose-6-phosphate; D019298:Glucose-6-Phosphate; D000135:Acid Phosphatase; D005952:Glucose-6-Phosphatase; D019826:Cholic Acid