Identification of rat liver glucose-3-phosphatase as an inositol monophosphatase inhibited by lithium.
Glucose-3-phosphatase (Glc3Pase) from rat liver has been purified 780-fold with a 4% recovery. The substrate specificity of the purified enzyme agreed with that of inositol monophosphatase (EC 3.1.3.25). D-Glucose 3-phosphate (D-Glc(3)P; K(m) = 200 microM) was hydrolyzed with an efficiency similar to DL-myo-inositol 1-monophosphate (DL-Ins(1)P; K(m) = 80 microM), since the ratio V(max)/K(m) was similar for both substrates. Purification data, coelution of activities, thermal inactivation curves, optimal pH, bivalent cation requirements, inhibition by Li+, molecular weight, and isoelectric pH comparisons supported that the hydrolysis of D-Glc(3)P and DL-Ins(1)P was catalyzed by a unique phosphohydrolase identified as a hepatic form of the lithium-sensitive inositol monophosphatase. That the hydrolysis of D-Glc(3)P is a genuine feature of inositol monophosphatases was confirmed because the enzyme purified from bovine brain showed also Glc3Pase activity, and inspection of published 3D models of inositol monophosphatase complexes with D(L)-Ins(1)P or D(L)-Ins(4)P indicated that beta(alpha)-D-Glc(3)P in a pyranose conformation with all (but one) the hydroxy groups in equatorial orientation would fit in the active site as other good substrates do. The results of this work are suggestive of possible relationships between inositol and sugar 3-phosphate metabolism.
Canales
J
J
Unidad de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Extremadura, Badajoz, Spain.
Buitrago
F
F
Faraldo
A
A
Avalos
M
M
Cameselle
J C
JC
D000818:Animals; D001921:Brain / Q000201:enzymology; D002417:Cattle; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008094:Lithium / Q000494:pharmacology*; D008099:Liver / Q000201:enzymology*; D008297:Male; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity
D008094:Lithium; C085852:glucose-3-phosphatase; D010744:Phosphoric Monoester Hydrolases; C021822:myo-inositol-1 (or 4)-monophosphatase