Characterization of polyisoprenyl phosphate phosphatase activity in rat liver.
The polyisoprenyl phosphate dephosphorylating activity of rat liver has been investigated with regard to substrate specificity, subcellular distribution, and transmembrane orientation. Total liver microsomes were employed as a source of enzymatic activity against a variety of 32P-labeled substrates. Susceptibility to dephosphorylation followed the order solanesyl phosphate greater than alpha-cis-polyprenyl 19-phosphate = alpha-trans-polyprenyl 19-phosphate = dihydrosolanesyl phosphate greater than (S)-dolichyl 19-phosphate = (R)-dolichyl 19-phosphate = (R,S)-dolichyl 11-phosphate. There appeared to be no major effect of chain length from 11 to 20 isoprenes. Data obtained from inhibition studies using solanesyl [32P]phosphate as substrate were consistent with the substrate specificity studies and suggested that a single activity is responsible. With dolichyl [32P]phosphate as substrate, the phosphatase specific activity of the subcellular fractions prepared from rat liver was found to follow the sequence Golgi = smooth endoplasmic reticulum greater than plasma membrane greater than lysosomes = rough endoplasmic reticulum greater than nuclei greater than mitochondria. Transmembrane topography studies, using enzyme latency as a criterion, were consistent with an orientation of the active site facing the cytoplasm.
D000818:Animals; D002451:Cell Compartmentation; D004288:Dolichol Phosphates / Q000378:metabolism; D004286:Dolichols / Q000378:metabolism; D007425:Intracellular Membranes / Q000201:enzymology / Q000648:ultrastructure; D008360:Mannosephosphates / Q000378:metabolism; D008862:Microsomes, Liver / Q000201:enzymology* / Q000648:ultrastructure; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D011106:Polyisoprenyl Phosphates / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity
D004288:Dolichol Phosphates; D004286:Dolichols; D008360:Mannosephosphates; D011106:Polyisoprenyl Phosphates; C048154:solanesyl phosphate; D010744:Phosphoric Monoester Hydrolases; C020088:dolichyl-phosphatase