Isolation of a Golgi apparatus-rich fraction from rat liver. IV. Thiamine pyrophosphatase.
The thiamine pyrophosphatase (the enzyme [s] catalyzing the release of inorganic phosphate with thiamine pyrophosphate as the substrate) activities of Golgi apparatus-, plasma membrane-, endoplasmic reticulum-, and mitochondria-rich fractions from rat liver were compared at pH 8. Activity was concentrated in the Golgi apparatus fractions, which, on a protein basis, had a specific activity six to eight times that of the total homogenates or purified endoplasmic reticulum fractions. However, only 1-3% of the total activity was recovered in the Golgi apparatus fractions under conditions where 30-50% of the UDPgalactose:N-acetylglucosamine-galactosyl transferase activity was recovered. Considering both recovery of galactosyl transferase and fraction purity, we estimate that approximately 10% of the total thiamine pyrophosphatase activity of the liver was localized within the Golgi apparatus, with a specific activity of about ten times that of the total homogenate. Cytochemically, reaction product was found in the cisternae of the endoplasmic reticulum as well as in the Golgi apparatus. This is in contrast to results obtained in most other tissues, where reaction product was restricted to the Golgi apparatus. Thus, enzymes of rat liver catalyzing the hydrolysis of thiamine pyrophosphate, although concentrated in the Golgi apparatus, are widely distributed among other cell components in this tissue.
D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D004721:Endoplasmic Reticulum / Q000201:enzymology; D006056:Golgi Apparatus / Q000201:enzymology*; D006651:Histocytochemistry; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology; D008110:Liver Extracts / Q000032:analysis*; D008297:Male; D008854:Microscopy, Electron; D008930:Mitochondria, Liver / Q000201:enzymology; D011755:Pyrophosphatases / Q000302:isolation & purification*; D051381:Rats; D013835:Thiamine Pyrophosphate / Q000378:metabolism*; D014166:Transferases / Q000032:analysis
D008110:Liver Extracts; D014166:Transferases; D011755:Pyrophosphatases; D013835:Thiamine Pyrophosphate