Relipidation of phospholipid-depleted microsomal particles with high glucose 6-phosphatase activity.
Microsomal particles enriched up to 20-fold in glucose 6-phosphatase activity (as compared to crude microsomal fractions) were prepared from livers of phenobarbital-treated, normal, and diabetic rats by a method involving sucrose-density gradient centrifugation through a layer containing deoxycholate, followed by flotation of the delipidated particles in a phospholipid-detergent mixture. The flotation with phospholipid resulted in the extraction of additional protein, in a corresponding increase in specific activity, and in relipidation of the particles to their original phospholipid content. Detergents alone did not extract any additional protein. Relipidation caused a change in the properties of the microsomal membrane, as indicated by a 4-fold decrease in the K(m) for glucose 6-phosphate and mannose 6-phosphate. The maximal rate was not affected. The crude homogenate of diabetic rat liver contained more latent enzyme than that of normal rats. The diabetic rats also yielded purified microsomal particles of specific glucose 6-phosphatase activity twice that of normal and five times that of phenobarbital-treated rats, indicating that some regulatory mechanism exists for the incorporation of different amounts of the enzyme into the endoplasmic reticulum.
D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D003840:Deoxycholic Acid; D003920:Diabetes Mellitus / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology; D005952:Glucose-6-Phosphatase / Q000378:metabolism*; D008099:Liver / Q000187:drug effects / Q000648:ultrastructure; D008862:Microsomes, Liver / Q000201:enzymology* / Q000378:metabolism; D010634:Phenobarbital / Q000494:pharmacology; D010743:Phospholipids / Q000378:metabolism*
D010743:Phospholipids; D003840:Deoxycholic Acid; D005952:Glucose-6-Phosphatase; D010634:Phenobarbital