Purification of a 32.5 kDa monomeric sulfotransferase from rat liver with activity for bile acids and phenolic steroids

AbstractBoth bile acid and phenolic steroid sulfotransferase activities in rat liver cytosol have previously been identified in fractions corresponding to apparent molecular masses of 60–70 and 30–35 kDa. We purified the latter activity corresponding to a monomeric protein. Activity for bile acids and phenolic steroids co-eluted on sequential chromatography on Sephadex G-75 sf, Affigel blue, chromatofocusing and hydroxyapatite. The protein was homogeneous on SDS-PAGE (32.5 kDa)

Interaction between dye and zinc in the dye-dispersing ZnO films prepared by a wet process

Dye-dispersing ZnO precursor gel films were prepared on indium tin oxide electrodes from a zinc acetate solution containing eosin Y by dip-coating, steam treatment, and then heating at a low temperature. The electronic interaction between the dye and zinc in the dye-dispersing gel films were investigated by spectroscopic and electrochemical measurements. A photocurrent was observed in the dye-dispersing gel electrodes before the steam treatment. The photocurrent value increased by the steam treatment and heating due to crystallization of the gel and removal of organic impurities. The dye molecules existed between the interlayers of the layered zinc hydroxide coexisting with the ZnO. The photoexcited electron in the dye should be injected into the ZnO conduction band via the layered zinc hydroxide. The value increased with an increase in the dye content even though the ZnO crystallinity decreased. The dye-zinc interaction, i.e., the complex formation and photoinduced electron injection, played an important role in the electron transport and photoelectric conversion.ArticleRESEARCH ON CHEMICAL INTERMEDIATES. 41(9):6559-6574 (2015)journal articl

Formation of ZnO thin films by photocatalytic reaction

Zinc oxide and layered zinc hydroxides were deposited from an aqueous solution of zinc nitrate at 323–358 K on a substrate plate with a very thin titanium dioxide film by a photocatalytic reaction. The amorphous or low crystalline zinc hydroxide aggregates were deposited at a low temperature. The zinc oxide crystals with about 1–2 μm-sized hexagonal columns and 10 nm-sized spheres were formed at 338–358 K. Nitrate ions in the solution were reduced to nitrite ions, and water was transformed into hydroxide ions by a photocatalytic reaction on the titanium dioxide film. The pH value increased on the substrate surface with the titanium dioxide film, which caused the zinc hydroxide formation on the film. The zinc hydroxides were then dehydrated and transformed into zinc oxide. The average crystallite size of the zinc oxide decreased with an increase in the reaction temperature because the reaction rates of the formation and dehydration of the zinc hydroxides increased which resulted in an increase in the formation rate of the crystal zinc oxide nuclei.ArticleAPPLIED CATALYSIS B-ENVIRONMENTAL. 160:651-657 (2014)journal articl

Involvement of SIK3 in Glucose and Lipid Homeostasis in Mice

Salt-inducible kinase 3 (SIK3), an AMP-activated protein kinase-related kinase, is induced in the murine liver after the consumption of a diet rich in fat, sucrose, and cholesterol. To examine whether SIK3 can modulate glucose and lipid metabolism in the liver, we analyzed phenotypes of SIK3-deficent mice. Sik3−/− mice have a malnourished the phenotype (i.e., lipodystrophy, hypolipidemia, hypoglycemia, and hyper-insulin sensitivity) accompanied by cholestasis and cholelithiasis. The hypoglycemic and hyper-insulin-sensitive phenotypes may be due to reduced energy storage, which is represented by the low expression levels of mRNA for components of the fatty acid synthesis pathways in the liver. The biliary disorders in Sik3−/− mice are associated with the dysregulation of gene expression programs that respond to nutritional stresses and are probably regulated by nuclear receptors. Retinoic acid plays a role in cholesterol and bile acid homeostasis, wheras ALDH1a which produces retinoic acid, is expressed at low levels in Sik3−/− mice. Lipid metabolism disorders in Sik3−/− mice are ameliorated by the treatment with 9-cis-retinoic acid. In conclusion, SIK3 is a novel energy regulator that modulates cholesterol and bile acid metabolism by coupling with retinoid metabolism, and may alter the size of energy storage in mice

1. Studies with the Purified Enzyme, Isolated Rat Hepatocytes, and Inhibition by Indomethacin

We recently identified that the Y ' bile acid binders are 3a-hydroxysteroid dehydrogenases (3a-HSD). In the present studies, purified 3a-HSD catalyzed rapid 3H loss from 13,1-3H, C24-4Cllithocholic and chenodeoxycholic acids without net conversion to 3-oxo bile acids under physiologic pH and redox conditions. 13,6-3HICholic acid was a poor substrate. The Y' fraction of hepatic cytosol was exclusively responsible for this activity and 3H was transferred selectively to NADP+. Timedependent 3H loss was also seen in isolated hepatocytes. Further hydroxylation products of lithocholic and chenodeoxycholic acids lost 3H at the same rate, whereas 3H loss from lithocholic acid rapidly ceased, which suggests compartmentation of this bile acid in hepatocytes. Indomethacin inhibited 3H loss from bile acids either in incubations with the pure enzyme or in isolated hepatocytes. Indomethacin did not alter the initial uptake rate of bile acids by hepatocytes, but caused a redistribution of unconjugated bile acids into the medium at early time points (2.5 and 5.0 min) and that of conjugated bile acids at later time intervals (30 min). 3H loss from the 3ft position therefore can be used to probe the interaction between bile acids and cytosolic 3a-HSD in intact cells, and indomethacin is capable of inhibiting this interaction