title: Localization of enzymes involved in glycero-ether bond formation in rat liver.
abstract: Studies on the localization of alkyldihydroxyacetone-phosphate synthase in rat liver are described. Less than 5% of the total activity was found in the cytosolic fraction, suggesting that the enzyme is membrane-bound. The ratio of the enzymatic (specific) activity over that of dihydroxyacetone-phosphate acyltransferase, a peroxisomal enzyme, is 10-fold higher in the microsomal fraction, when compared to the peroxisomal fraction. Studying the distribution of the enzyme in a linear density gradient, two activity peaks were found in the peroxisomal and the microsomal fraction indicating a bimodal localization of alkyldihydroxyacetone-phosphate synthase. Rabert et al. (Rabert, U., Völkl, A. and Debuch, H. (1986) Biol. Chem. Hoppe-Seyler 367, 215-222) have presented evidence that the activity in the microsomal fraction was mainly caused by a different enzyme that preferentially converted acyldihydroxyacetone to alkyldihydroxyacetone. We also found a radioactive product, different from alkyldihydroxyacetone-phosphate, upon incubation of microsomal protein in the presence of [14C]hexadecanol. However, it was shown that this product was formed independently of the presence of acyldihydroxyacetone. The product yielded [14C]hexadecanol upon alkaline hydrolysis, clearly showing that it did not contain an ether-bond. Upon incubation of microsomal protein with [14C]palmitic acid and hexadecanol the product was also observed and its chromatographic behaviour resembled that of a synthetically prepared palmitoyl ester of hexadecanol. From these data it was concluded that the product formed is most likely a wax and that the enzyme responsible for this conversion is clearly different from the alkyldihydroxyacetone-phosphate synthase. The implication is that acyldihydroxyacetone-phosphate cannot be replaced by acyldihydroxyacetone in the process of glycero-ether bond formation.
authors.affiliation: Centre for Biomembranes and Lipid Enzymology, University of Utrecht, The Netherlands.
authors.forename: D
authors.initials: D
authors.lastname: Hardeman
authors.forename: H
authors.initials: H
authors.lastname: van den Bosch
mesh_terms: D019883:Alkyl and Aryl Transferases; D000818:Animals; D002458:Cell Fractionation; D002855:Chromatography, Thin Layer; D004987:Ethers / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008297:Male; D008830:Microbodies / Q000201:enzymology*; D008862:Microsomes, Liver / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D014166:Transferases / Q000378:metabolism*
chemical_list: D004987:Ethers; D014166:Transferases; D019883:Alkyl and Aryl Transferases; C019654:alkylglycerone-phosphate synthase