Reversible thermal inactivation of the quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus. Ca2+ ions are necessary for re-activation.
The soluble form of the homogeneous quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus is reversibly inactivated at temperatures above 35 degrees C. An equilibrium is established between active and denatured enzyme, this depending on the protein concentration and the inactivation temperature used. Upon thermal inactivation the enzyme dissociates into the prosthetic group pyrroloquinoline quinone and the apo form of glucose dehydrogenase. After inactivation at 50 degrees C active enzyme is re-formed again at 25 degrees C. Ca2+ ions are necessary for the re-activation process. The velocity of re-activation depends on the protein concentration, the concentration of the prosthetic group pyrroloquinoline quinone and the Ca2+ concentration. The apo form of glucose dehydrogenase can be isolated, and in the presence of pyrroloquinoline quinone and Ca2+ active holoenzyme is formed. Even though native glucose dehydrogenase is not inactivated in the presence of EDTA or trans-1,2-diaminocyclohexane-NNN'NH-tetra-acetic acid, Ca2+ stabilizes the enzyme against thermal inactivation. Two Ca2+ ions are found per subunit of glucose dehydrogenase. The data suggest that pyrroloquinoline quinone is bound at the active site via a Ca2+ bridge. Mn2+ and Cd2+ can replace Ca2+ in the re-activation mixture.
Geiger
O
O
Institut für Mikrobiologie der Universität Hohenheim, Federal Republic of Germany.
Görisch
H
H
D000150:Acinetobacter / Q000201:enzymology*; D001051:Apoenzymes / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology*; D002237:Carbohydrate Dehydrogenases / Q000037:antagonists & inhibitors*; D003067:Coenzymes / Q000494:pharmacology; D004793:Enzyme Reactivators / Q000494:pharmacology*; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006358:Hot Temperature; D045542:PQQ Cofactor; D015363:Quinolones / Q000494:pharmacology
D001051:Apoenzymes; D003067:Coenzymes; D004793:Enzyme Reactivators; D015363:Quinolones; D045542:PQQ Cofactor; D002237:Carbohydrate Dehydrogenases; D005948:Glucose Dehydrogenases; D042843:Glucose 1-Dehydrogenase; D002118:Calcium