Regulation of function and protein expression of noradrenaline transporter by protein kinases
| Project/Area Number |
11680763
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurochemistry/Neuropharmacology
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| Research Institution | Showa University |
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Principal Investigator |
KIUCHI Yuji Showa University, School of Pharmaceutical Sciences, Professor, 薬学部, 教授 (50204821)
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| Co-Investigator(Kenkyū-buntansha) |
OYAMADA Hideto Showa University, School of Medicine, Instructor, 医学部, 助手 (50266160)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Noradrenaline transporter / Noradrenaline reuptake / Ca^<2+> / calmodulin-dependent kinase II / Protein phosphorylation / Intracellular translocation / Regulation of expression / PC12 cells / Green fluorescent protein (GFP) / 遺伝子発現 / トランスロケーション |
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| Research Abstract |
Regulation of noradrenaline transporter (NAT) function by calmodulin-related protein kinases was studied in PC12 cells, which endogenously express NA transporters. 1. We determined a full sequence of rat NAT cDNA and found five consensus sites for phosphorylation by Ca^<2+>/calmodulin-dependent (CaM) kinase II.A synthetic peptide whose sequence was contained in the intracellular COOH-terminal domain of rat NAT was phosphorylated by purified brain CaM kinase II.2. Extensive NAT-like immunoreactivity or fluorescence of transfected GFP-NAT fusion protein in plasma membrane of PC12 cells disappeared after incubation with KN-93 or in the absence of Ca^<2+>. 3. In clonal cell lines of PC12 cells which overexpressed CaM kinase II α subunit, [^3H]NA uptake as well as expression of NAT mRNA and protein was markedly increased compared with wild type cells. Content of phosphorylated CREB, an active form of a transcription factor was also increased in CaM kinase II α-overexpressed cell lines. The increased uptake and mRNA expression were significantly suppressed by chronic treatment of KN-93. These results suggest that NA uptake might be facilitated by Ca^<2+>/calmodulin-dependent protein kinases through direct phosphorylation of COOH-terminal domain of NAT and/or through stimulated translocation of NAT to plasma membrane, and that activation of CaM kinase II might chronically enhance expression of NAT.
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Report
(2 results)
Research Products
(7 results)
