Demonstration of glyoxalase II in rat liver mitochondria. Partial purification and occurrence in multiple forms.
Glyoxalase II (S-(2-hydroxyacyl)glutathione hydrolase, EC 3.1.2.6), which has been regarded as a cytosolic enzyme, was also found in rat liver mitochondria. The mitochondrial fraction contained about 10-15% of the total glyoxalase II activity in liver. The actual existence of the specific mitochondrial glyoxalase II was verified by showing that all of the activity of the crude mitochondrial pellet was still present in purified mitochondria prepared in a Ficoll gradient. Subfractionation of the mitochondria by digitonin treatment showed that 56% of the activity resided in the mitochondrial matrix and 19% in the intermembrane space. Partial purification of the enzyme (420-fold) was also achieved. Statistically significant differences were found in the substrate specificities of the mitochondrial and the cytosolic glyoxalase II. Electrophoresis and isoelectric focusing of either the crude mitochondrial extract or of the purified mitochondrial glyoxalase II resolved the enzyme activity into five forms with the respective pI values of 8.1, 7.5, 7.0, 6.85 and 6.6. Three of these forms (pI values 7.0-6.6) were exclusively mitochondrial, with no counterpart in the cytosol. The relative molecular mass of the partially purified enzyme, as estimated by Superose 12 gel chromatography, was 21,000. These results give evidence for the presence of mitochondrial glyoxalase II which is different from the cytosolic enzymes in several characteristics.
Talesa
V
V
Department of Medical Chemistry, University of Helsinki, Finland.
Uotila
L
L
Koivusalo
M
M
Principato
G
G
Giovannini
E
E
Rosi
G
G
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D003600:Cytosol / Q000201:enzymology; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology*; D008970:Molecular Weight; D051381:Rats; D013379:Substrate Specificity; D013869:Thiolester Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D000099301:Hydroxyacylglutathione Hydrolase
D007527:Isoenzymes; D013869:Thiolester Hydrolases; D000099301:Hydroxyacylglutathione Hydrolase