Subcellular distribution and properties of carbonyl reductase in guinea pig lung.
On subcellular fractionation, carbonyl reductase (EC 1.1.1.184) activity in guinea pig lung was found in the mitochondrial, microsomal, and cytosolic fractions; the specific activity in the mitochondrial fraction was more than five times higher than those in the microsomal and cytosolic fractions. Further separation of the mitochondrial fraction on a sucrose gradient revealed that about half of the reductase activity is localized in mitochondria and one-third in a peroxidase-rich fraction. Although carbonyl reductase in both the mitochondrial and microsomal fractions was solubilized effectively by mixing with 1% Triton X-100 and 1 M KCl, the enzyme activity in the mitochondrial fraction was more highly enhanced by the solubilization than was that in the microsomal fraction. Carbonyl reductases were purified to homogeneity from the mitochondrial, microsomal, and cytosolic fractions. The three enzymes were almost identical in catalytic, structural, and immunological properties. Carbonyl reductase, synthesized in a rabbit reticulocyte lysate cell-free system, was apparently the same in molecular size as the subunit of the mature enzyme purified from cytosol. These results indicate that the same enzyme species is localized in the three different subcellular compartments of lung.
D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D007700:Kinetics; D008168:Lung / Q000201:enzymology* / Q000648:ultrastructure; D008861:Microsomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D008970:Molecular Weight; D009249:NADP / Q000494:pharmacology; D010446:Peptide Fragments / Q000302:isolation & purification; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D012697:Serine Endopeptidases; D012995:Solubility; D013347:Subcellular Fractions / Q000201:enzymology*
D010446:Peptide Fragments; D012333:RNA, Messenger; D009249:NADP; D000429:Alcohol Oxidoreductases; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase