| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1995: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1994: ¥5,400,000 (Direct Cost: ¥5,400,000)
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| Research Abstract |
1. Rabbit anti-Na / myo-inositol cotransporter (SMIT) antibody were raised against the synthetic 51 amino acid peptide corresponding to the region which had no similarity to any proteins reporte previously.
2. Anti-SMIT antibody detected 4 bands in rat kidneys in immunoblotting analyzes. Their molecular sizes were 76-, 66-, 57- and 47- kDa, and the 76- kDa band was very faint ; much smaller than the value, 79.5kDa, calculated from the cDNA.The 66- and 47- dDa band were observed in the mitochondrial and cytosolic fractions, and 57- kDa band was limited to the basolateral membrane fraction.
3. Rabbit antisera against renal water channels (WCH-CD,MIWC,AQP3) and Cl-channels (CIC-K1, pIC1n) were raised against synthetic peptides corresponding to the parts of their amino acid sequences.
4. The size of the detected WCH-CD,MIWC and AQP3 were 47-, 46- and 31-kDa. The former 2 values were significantly larger than those from their cDNA and the remaing was coincided with the value from cDNA.All bands relating to 3 water channels were only observed in the mitochondria fraction, and not in thebasolateral membrane fraction.
5. The anti-CIC-K1 and anti-pIC1n sera detected 36-and 50-kDa bands, of which values were much different to those calculated from their cDNAs. The ClC-K1-band was widespread : cytosol, mytochondria and basolateral membranes. The pIC1n-band was detected in the mitochondria fraction, and very slightly in the basolateral membrane fraction.
6. The present results show the great or extensive processing after translation from the mRNAs of the transporeter and channels. Their presence in mitochondria also has been worthy of remark ; the possibility of response of mitochondria to cytoplasmic osmotic pressure or concenetrations of ions
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