To characterize the sialyltransferase-IV activity in brain tissues, the activities of GMl b-, GD1 a-, GT1 b-, and GQl c-synthases in adult cichlid fish and rat brains were examined using GA1, GM1, GD1 b, or a cod brain ganglioside mixture as the substrate. The GD1 a-synthase activity in the total membrane fraction from cichlid fish brain required divalent cations such as Mg2~o r Mn 2' and Triton CF-54 for its full activity. The Vmax value was 1,340 pmol/mg of protein/h at an optimal pH of 6.5, whereas the apparent Km values for CMP-sialic acid and GM1 were 172 and 78 NM, respectively . Cichlid fish and rat brains also contained GM1 b-, GT1 b-, and GQ1 c-synthase activities. The ratio of GM1 b-, GD1 a-, and GT1 b-synthase activities in fish brain was 1 .00 :0 .89:1 .13, respectively, and in rat brain 1 .00:0 .60:0 .63. Incubation of fish brain membranes with a cod brain ganglioside mixture, which contains GT1c, and [3H]CMP-sialic acid produced radiolabeled GQ1 c. It is interesting that the adult rat brain also contains an appreciable level of GQ1 c-synthase activity despite its very low concentrations of c-series gangliosides. The GD1 a-or GQl c-synthase activity in fish and rat brain was inhibited specifically by coincubation with the glycolipids that serve as the substrates for other sialyltransferase-IV reactions. Thus, the GD1 a-synthase activity was inhibited by GA1 and GD1 b, but not by LacCer, GM3, or GD3 . In a similar manner, the synthesis of GQ1c was suppressed by GA1, GM1, and GD1b, but not by LacCer, GM3, or GD3. The GDla-synthase activity directed toward endogenous GM1 was inhibited by GA1 or GT1 b, whereas the endogenous GT1 b-synthase activity was suppressed by GA1 or GM1 . GA1, GM1, and GD1 b did not affect the endogenous GM3-and GD3synthase activities . These results clearly demonstrate that sialyltransferase-IV in brain tissues catalyzes the reaction for GQ1 c synthesis in the c-pathway as well as the corresponding steps in the asialo-, a-, and b-pathway in ganglioside biosynthesis . Key Words: Sialyltransferase-Gangliosides-GQ1c-Brain -Fish-c-Series gangliosides. J. Neurochem. 64, 385-393 (1995) .Gangliosides are sialic acid-containing glycosphingolipids and are enriched especially in vertebrate ner-nomenclature is according to Svennerholm (1963) .