| Project/Area Number |
06454648
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Functional biochemistry
|
|---|
| Research Institution | HOKKAIDO UNIVERSITY |
|---|
Principal Investigator |
TANIGUCHI Kazuya HOKKAIDO UNIV., GRADUATE SCHOOL OF SCIENCE,PROFESSOR, 大学院・理学研究科, 教授 (40028204)
|
|---|
| Project Period (FY) |
1994 – 1996
|
| Project Status |
Completed (Fiscal Year 1996)
|
| Budget Amount *help |
¥7,300,000 (Direct Cost: ¥7,300,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1995: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1994: ¥4,700,000 (Direct Cost: ¥4,700,000)
|
|---|
| Keywords | Regulation / Na, K-ATPase / H,K-ATPase / Phosphorylation / キナーゼ / 胃壁H^+ポンプ / ATPase / 脱リン酸化 / カルシウムイオン / Na^+ポンプ / P型ポンプ / エネルギー変換 / ナトリウムポンプ / H+ポンプ / 構造変化 / タンパクキナーゼ / フォスファターゼ / チロシンキナーゼ / Na^+,K^+-ATPase / H^+,K^+-ATPase / イオン輸送ポンプ / プロトンポンプ |
|---|
| Research Abstract |
When pig stomach membrance H,K-ATPase preparations were incubated with ^^<32>ATP,Mg^<2+> and Ca^<2+>, reversible phosphorylation of specific Tyr and Ser residues in the N-terminal alpha-chain of H,K-ATPase occurred without any detectable phosphorylation in other regions of the chain. Amino acid sequence analysis of purified peptides and others showed a sequential phosphorylation of Tyr-10 and Tyr-7 and sub muM Ca^<2+> activated phosphorylation of Ser-27. The apparent molecular weight of a detergent solubilized Tyr-kinase was estimated to be around 50 kDa by a gel filtration column and the kinase activity detected on a gel band after renaturation. The Ser-kinase present was suggested to be a conventional PKC.H,K-ATPase preparations phosphorylated fusion proteins consised of maltose binding protein and a peptide portion of the alpha-chain from Gly-2 to Gln-111. These data and others suggested that N-terminal cytosolic domain is sufficient for phosphorylation by endogenous Tyr-kinase and PKC in the membrane H,K-ATPase preparations.
|
