1999 Fiscal Year Final Research Report Summary
The function and expression-mechanism of growth-associated protein GAP-43 in neuron
| Project/Area Number |
10680728
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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 | Fukuok University |
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Principal Investigator |
TAKAMI Noboru Fukuoka Univ., Radioisotope Labo. Sch. Med. Assist. Prof., 医学部・RI施設, 助手 (80154904)
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| Project Period (FY) |
1998 – 1999
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| Keywords | GAP-43 / synaptic vesicle / exocytosis |
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| Research Abstract |
GAP-43 is a nerve-specific protein that is a predominantly localized in the axonal growth cones and presynaptic terminal membrane, but its precise function is nuclear, GAP-43 is attached to the presynaptic membrane by two acylated Cys at positions 3 and 4. The localization of wild and mutated (C3C4→A3A4 and C3→A3) GAP-43 was examined by cell-fractionation and immunofluorescence staining after transfection with each plasmid in COS-1 or PC12 cells. The mutant (C3C4→A3A4) was mainly recovered in the cytoplasmic fraction and fraction and stained at the neurites. In contrast, the mutant (C3→A3) was detected in both of them. Therefore it was shown that the fatty acylation of these Cys is essential for targeting GAP-43 to the growth cones or presynaptic membrane. Moreover, to identify the components interacted with GAP-43 I have used the yeast two-hybrid system. Then I obtained calmodulin and myosin light-chain in this screen and confirmed as the positive clones by retransformation. In addition, the experiments in vitro revealed that GAP-43 interacted p115/TAP which is implicated in the intracellular vesicular transcytosis. Taken together, these results suggested that GAP-43 is participated in the regulation of synaptic vesicle exocytosis with neurotransmitter release at the active zone of presynaptic membrane, since we have reported the interaction of GAP-43 with SNARE complex.
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[Publications] Arima, K., Nkamura, Y., Takami, N., Mizusaki, K., Yoshida, N. and Okamoto, K.: "AnィイD2α2ィエD2-macroglobulin-serine proteinnase complex from human carcinomatous ascites and pleural effusion : monoclonal antibody preparation, and immunohistochemical study"Oncology Res.. 10. 499-507 (1998)
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