1992 Fiscal Year Final Research Report Summary
Analysis of ganglioside-dependent protein phosphorylation in rat brain membrane
| Project/Area Number |
03680139
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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 |
物質生物化学
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| Research Institution | The Institute of Physical and Chemical Research(RIKEN) (1992)
The University of Tokyo (1991) |
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Principal Investigator |
TSUJI Shuichi The Institute of Physical and Chemical Research(RIKEN) Frontier Research Program Team Reader, 国際フロンティア研究システム, チームリーダー (90124677)
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| Project Period (FY) |
1991 – 1992
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| Keywords | Ganglioside / Protein phosphorylation / Protein kinase / Phosphoprotein phosphatase |
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| Research Abstract |
1)Whether or not a ganglioside induces the protein phosphorylation in the rat brain membrane fraction was investigated. Phosphorylation of the 72KDa protein was significantly affected by the addition of 80nN GQ1b in vitro, which is far below the reported concentration of gangliosides that affects protein phosphorylation in the neuronal membrane fraction. This action of GQ1b was bimodal : it being not only stimulatory as to the incorporation of phosphate into the 72KDa protein on incubation for 20 sec, but also as to the release of phosphate from or breakdown of the 72KDa protein on incubation for more than 5 min. Eighty nM GQ1b did not noticeably affect ATPase in the same fraction. These results suggest that the transphosphorylation of the 72KDa protein affected by the interaction of GQ1b with either the responsible enzymes or the 72KDa protein as a substrate.
2)72KDa protein was successively solvilized and purified to the homoginous state so far seen on SDS-PAGE, using DEAE-Sepharose, Octyl-Sepharose and Heparin-Sepharose Chromatography, followed by gel filtration with G3000SW. After sequencing of the lysylendopeptidase-fragments of 72KDa protein, several cDNAs encoding the full length of 72KDa protein were cloned.
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