| Co-Investigator(Kenkyū-buntansha) |
FUKAMI Yasuo Kobe University, Research Center for, Molecular Biology Associate Professor, 遺伝子実験施設, 助教授 (00156746)
TACHIBANA Hideki Kobe University, Faculty of Science, Associate Professor, 理学部, 助教授 (70126118)
KISHIMOTO Akira Kobe University, School of Medicine, Associate Professor, 医学部, 助教授 (60127363)
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| Research Abstract |
Recent analysis has revealed that protein kinase C is a large family, with the multiple subspecies that possess subtle individual enzymological characteristics and different distribution in particular cell types. Presumably, each member of the family plays discrete roles in the processing of various physiological and responses to external signals. For identifying the specific function of each subspecies of protein kinase C, we focused on the mechanism of release response, growth control and differentiation.
(1) Release response-synaptosome Synaptosomes prepared from hippocampus contains less gamma PKC activity compared with beta and alpha PKC. We could succeeded in preparing the synaptosomes containing only gamma PKC by pre-treatment of TPA, which induced selective down-regulation of beta and alpha PKC. Using these synaptosomes we analyzed the involvement of gamma PKC in noradrenalin (NA) release response. No significant difference of NA release induced by TPA was observed between the s
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ynaptosomes containing both beta and alpha PKC and only gamma PKC. This result indicated that gamma PKC did not involve in the release response.
(2) Growth control-S. cerevisiae In contrast to the establishment of the role of cyclic AMP dependent protein kinase in the regulation of yeast cell cycle, no experimental evidence about the existence of PKC has been reported. We could detect multiple subspecies of PKC also in S. cerevisiae, which were activated by the simultaneous addition of Ca^<2+>, diacylglycerol, and phosphatidylserine. We have started to isolate the genes of PKC from S. cerevisiae, which may provide clues to elucidate definite functions of PKC in the regulatory role of growth and cell cycle.
(3) Differentiation-HL-60 It is well established that TPA and retinoic acid (RA) can induce HL-60 cell to differentiate toward granulocyle. Analysis of PKC-subspecies reveals that in addition to alpha and beta PKC, unidentified subspecies is expressed in HL-60 cell. The expression of these three subspecies were differently regulated during the differentiation toward granulocyte. Less
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