| Project/Area Number |
61571056
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Biological pharmacy
|
|---|
| Research Institution | Faculty of Pharmaceutical Science, University of Shizuoka (1987)
Shizuoka College of Pharmacy (1986) |
|---|
Principal Investigator |
HIRABAYAHSI Yoshio Department of Biochemistry, Faculty of Pharmaceutical Science, University of Shizuoka, 薬学部, 助手 (90106435)
|
|---|
| Project Period (FY) |
1986 – 1987
|
| Project Status |
Completed (Fiscal Year 1987)
|
| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1987: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1986: ¥1,400,000 (Direct Cost: ¥1,400,000)
|
|---|
| Keywords | Melanoma antigen / sialic acid / gangliosides / ガングリオシド / シアル酸転移酵素 / シアリルトラントフェラーゼ |
|---|
| Research Abstract |
The immunological and chemical properties of melanoma antigens have been analyzed by using C57BL/6 derived B16 melanoma cells. We have established two syngeneic monoclonal antibodies against the melanoma antigens. One monoclonal antibody, M2590, recognized the interspecies cross-reactive melanoma antigenic determinant widely shared in various mammalian species. This antibody was shown to have binding capacity for GM3(NeuAc) ganglioside. GM3 is known to be distributed in various cell types including human red blood cells. M2590, however, does not necessarily react with all types of cells having GM3 ganglioside. The reactivity of GM3 with cells seems largely to depend on the concentration of GM3 on the cell surface. In fact, B16 melanoma cells have strong enzymic ativity to form GM3 ganglioside from lactosylceramide, when compared to normal cells. To understand the molecular and genetic mechanisms of the expression of GM3, it is essential to purify an enzyme involved in teh synthesis of GM3 in B16 melanoma cells. The sialyltransferase is a membrane bound enzyme and can be solubilized by using 1% triton X-100. The extract was applied to hydroxyapatite/Q-Sepharose column. The fraction that passed through the column was adsorbed on an affinity column of lactosylceramide acid-Sepharose. The affinity purified enzyme have Mr 46000 by SDS-PAGE analysis. We are trying further and to determine the amino acid sequence of the molecule.
|
