| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
1. We isolated mitochondrial (mt) nuclei from anaerobically-grown cells, respiratory deficient cells and respiration-inhibited cells. We found that these mt-nuclei contained significantly reduced amount of 67 kDa protein, compared with mt-nuclei from aerobically-grown cells.
2. The N-terminal amino acid sequence of the 67 kDa protein was determined. This protein was proved to be a carnitine acetyltransferase (CAT) that localized in mitochondria. Immunological assay using the antibody against CAT suggested association of CAT with mt-nuclei.
3. Behavior of mitochondria, mt-nuclei, microtubules and actin were investigated by fluorescence microscopy in the triangular yeast, Trigonopsis variabilis.
4. We demonstrated that Abf2p, a major DNA-binding protein of mitochondria, is tightly associated with mt-nuclei by Western blotting and immunofluorescence microscopy. We showed that SDS-DNA PAGE is an efficient for detection of Abf2p on gels.
5. We identified the YMN-1 antigen protein of mt-nuclei, which is recognized by YMN-1 monoclonal antibody, as dihydrolipoyl transsuccinylase (KE2). Enzyme assay suggested that α-ketoglutarate dehydrogenase complex (KGDC) cosedimented with mt-nuclei in sucrose-density gradient.
6. Protein components of mt-nuclei were separated by two-dimensional (2D) electrophoresis. We determined N-terminal amino acid sequence of 54 kDa, 52 kDa, 50 kDa, 38 kDa and 22 kDa proteins, respectively, which are involved in mt-nuclei.
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