| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2009: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2008: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Research Abstract |
We prepared nuclear envelopes (NEs) from mouse pancreatic acinar cells and observed MaxiK currents in the membrane subjected to the patch-clamp technique. The results indicate that the MaxiK channels are oriented in the ER membrane as their usual extracellular side faces to cytosol. These observation is inconsistent with the conventional understandings of the protein transport process. In this study, we examined the orientation of MaxiK channel in the ER membrane of the pancreatic acinar cells.
We performed an immunofluorescent staining in our NE preparation using anti-Calnexin antibodies. Calnexin, an ER resident membrane protein, direscts C-terminus to the cytosol and N-terminus to the lumen. The antibody which binds to N-terminus of Calnexin showed the signal only when NE membrane was permeablilized. It supports the notion that the orientaion of our NE preparation itself would be intact. Collectively, we suggest that MaxiK channels of the mouse pancreatic acinar cells have unconventional orientation in the ER membrane.
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