Characterization and developmental expression of a novel sulfotransferase for the biosynthesis of sulfoglucuronyl glycolipids in the nervous system.
Sulfoglucuronyl glycolipids (SGGLs) are temporally and spatially regulated molecules in the developing nervous system. A novel sulfotransferase (ST) from rat brain which catalyzes the terminal step in the biosynthesis in vitro of SGGLs is described. The enzyme catalyzes a transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate to a hydroxyl group on carbon 3 of the terminal glucuronyl residue in IV3 beta-glucuronyl neolactotetraosylceramide (GlcAnLcOse4Cer) and VI3 beta-glucuronyl neolactohexaosylceramide (GlcAnLcOse6Cer) to form 3-sulfated glucuronyl glycolipids. The enzyme is highly specific for glucuronylglycolipids (GGLs) and requires the free-COOH group of the terminal glucuronic acid for reactivity. GGL:ST present in the microsomal membranes requires Mn2+ ions and a nonionic detergent, Triton X-100 for activity. The optimal pH is 7.2 with Tris-HCl buffer and Km values were 7 microM for 3'-phosphoadenosine 5'-phosphosulfate and 29 microM for GlcAnLcOse4Cer. GGL:ST was shown to be different from previously well studied galactocerebroside:sulfotransferase for the synthesis of myelin membrane-specific lipid sulfatide. This conclusion was based upon several criteria, i.e. including different requirements of incubation conditions for maximal activity, substrate competition experiments, different effects of heat, dithiothreitol, NaCl, and pyridoxal phosphate, as well as different profiles of expression of activity during development of the nervous tissues. The two enzymes were also partially resolved on a pyridoxal phosphate-ligated agarose column. Studies on the developmental expression of the GGL:ST in the rat cerebral cortex and cerebellum showed that it is not a regulatory enzyme controlling the expression of SGGLs in these neural tissues.
Chou
D K
DK
Department of Biochemistry, Eunice Kennedy Shriver Center for Mental Retardation, Waltham, Massachusetts 02254.
Jungalwala
F B
FB
D000375:Aging / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000201:enzymology* / Q000254:growth & development; D002240:Carbohydrate Sequence; D002531:Cerebellum / Q000201:enzymology; D002540:Cerebral Cortex / Q000201:enzymology; D002846:Chromatography, Affinity; D005260:Female; D005915:Globosides / Q000096:biosynthesis*; D006017:Glycolipids / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology*; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D011247:Pregnancy; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000302:isolation & purification / Q000378:metabolism*; D013466:Sulfurtransferases / Q000302:isolation & purification / Q000378:metabolism*
D005915:Globosides; D006017:Glycolipids; C063678:3-sulfoglucuronyl-lacto-N-neotetraosylceramide; C063679:3-sulfoglucuronyl-lacto-N-norhexaosylceramide; D011732:Pyridoxal Phosphate; D013466:Sulfurtransferases; C078859:IV(3)beta-glucuronyl neolactotetraosylceramide sulfotransferase; D015238:Sulfotransferases; C020556:galactosylceramide sulfotransferase