1995 Fiscal Year Final Research Report Summary
Protein-carbohydrate interactions : recognition and neural function of the mollusc (Aplysia) lectin.
| Project/Area Number |
06680755
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurochemistry/Neuropharmacology
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| Research Institution | Niigata University |
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Principal Investigator |
ABE Sachiko Niigata University, Brain Research Institute, Assistant, 脳研究所, 助手 (60018603)
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| Co-Investigator(Kenkyū-buntansha) |
ODANI Shoji Niigata University, Faculty of Science, Professor, 理学部, 教授 (60018702)
ARAKI Shigeko Niigata University, Brain Research Institute, Assistant, 脳研究所, 助手 (70018604)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Aplysia nervous tissue / Glycolipid binding protein / Phosphonoglycosphingo-lipid / Nerve bundle-specific |
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| Research Abstract |
Since we found a new family glycosphingolipids, phosphonoglycosphingolipids, in Aplysia kurodai, the structures and the localizations of these glycolipids have been investigated. Antiserum against FGL-IIb, which is specifically concentrated in the nerve fibers of Aplysia, stained only nerve bundles of Aplysia. So, it was considered that FGL-IIb may recognize the some components on the extracellular matrix. Following results for 2 years were obtained.
1. In 1994, Aplysia nervous tissues were solubilized by following three detergents, cholic acid sodium salt, Triton X-100 and octylglucoside. When the solubility of proteins and the recovery of glycolipids of Aplysia nervous tissues were investigated using above detergents, octylglucoside was best solubilizer in this experiment. Solubilized material of Aplysia nervous tissues treated with 100 mM octylglucoside was applied to anti-FGL-IIb antibody column and the column was washed with 50 mM octylgucoside and then eluted with 0.1% SDS solution. Silver staining by SDS-PAGE analysis of the 0.1% SDS eluate stained 45 kDa band more strongly than normal IgG column eluate.
2. In 1995, glycolipid fraction that was extracted with chloroform/methanol (2 : 1) from the 0.1% SDS eluate fraction was positively dot immunostained only by anti-FGL-IIb antiserum, but antisera against other glycolipids were not reacted. Coomassie Blue staining by SDS-PAGE of the another 0.1% SDS eluate fraction stained positively 45 kDa band.
Therefore, we reported that FGL-IIb which specifically localized in Aplysia nerve bundles formed the complex with 45 kDa protein in the presence of Ca^<2+> on the extracellular matrix. Study on the primary sequence of the 45 kDa protein is in progress.
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