| Project/Area Number |
61440089
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
物質生物化学
|
|---|
| Research Institution | Hokkaido University |
|---|
Principal Investigator |
MAKITA Akira Hokkaido University School of Medicine, 医学部, 教授 (60004561)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KASI Noriyuki Hokkaido University School of Medicine, 医学部, 助教授 (60001947)
HONKE Kouichi Hokkaido University School of Medicine, 医学部, 助手 (80190263)
GASA Shinsei Hokkaido University School of Medicine, 医学部, 助教授 (10142712)
|
|---|
| Project Period (FY) |
1986 – 1987
|
| Project Status |
Completed (Fiscal Year 1987)
|
| Budget Amount *help |
¥29,500,000 (Direct Cost: ¥29,500,000)
Fiscal Year 1987: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 1986: ¥25,000,000 (Direct Cost: ¥25,000,000)
|
|---|
| Keywords | Ganglioside GM1 synthase / GM3 synthase / GD3 synthase / Galactosyltransferase / Sialyltransferase / シアリルトランスフェラーゼ / スルフォトランスフェラーゼ / (β1→4)N-アセチルガラクトサミン転移酵素 / ガングリオシド【GM_2】 / (β1→3)ガラクトース転移酵素 / ガングリオシド【GM_1】 / ラット肝 / シアリルラクトース |
|---|
| Research Abstract |
1. GM_2 synthase which catalyzes the conversion of ganglioside GM_3 to GM_2 was purified over 6300-fold from rat liver particulate fractions by varius chromatographies. The purified enzyme consisted of two identical subunits of 64,000 dalton. GM_2 containing either NeuAc or NeuGc and the oligosaccharide, sialyllactose were good substrates for GM_3 synthase, indicating that the preferred acceptor has a structure ofNeuAc(or NeuGc) 2-3GalBl-4Glc-OR.
2. GM_1 synthase which forms GM_1 from GM_2 was purified from rat liver Golgi apparatus by affinity chromatographies to a 10,2000-fold. The synthase acted partly (28% of GM_2) on Gg_3Cer, but not on other glycolipids and glycoproteins tested.
3. We have developed a new method for assay of GM_3 aynthase, which forms GM_3 from LacCer, using a DEAE-Sephadex column. Using this method, GM_3 synthase was partialy purified from particulate fraction of pig spleen by conventinal and affinity chromatographies.
4. GD_3 synthase which forms GD_3 from GM_3 was fractionated from rat liver Golgi apparatus by conventional and affinity chromatographies. Factors and conditions affecting the synthase were determined.
5. Cerebroside sulfotransferase was purified from rat kidney, in parallel with the above studies. The purified enzyme had a wide range of substrate specificity acted on LacCer (110%) and Gg_3Cer (85%) in addition to Gal cer (100%), indicating that the same enzyme sulfates these glycolipids.
|
