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- W1551333215 abstract "This chapter describes the purification method for HA-tagged Rabphilin- 3 from the membrane fraction of overexpressing Spodoptera frugiperda cells (Sf9 cells). The chapter then describes the methods for detecting the interactions of Rabphilin-3 with α-actinin and rabaptin5, and the effects of Rabphilin-3 on the α-actinin-induced actin filament bundling and the receptor-mediated endocytosis. The steps used in the purification of HA-tagged Rabphilin-3 are as follows: (1) preparation of the membrane fraction from Sf9 cells, (2) heparin-Sepharose CL-6B column chromatography, and (3) Superose 12 HR10/30 column chromatography. The HA-tagged N-terminal fragment (amino acids 1–280) and HAtagged C-terminal fragment (amino acids 396–704) of Rabphilin-3 are purified by the same procedures as described above except that the Sf9 cells expressing each fragment are used. It has been shown that abnormalities of synaptic transmission and synaptic plasticity, which are observed in Rab3A-deficient mice, are not observed in Rabphilin-3-deficient mice. However, this does not necessarily indicate that Rabphilin-3 is not involved in neurotransmitter release. It is possible that Rim, another target molecule for Rab3A, compensates for the loss of function of Rabphilin-3 in the mice that abnormalities of synaptic transmission and synaptic plasticity, which are still not detected by the methods performed." @default.
- W1551333215 created "2016-06-24" @default.
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- W1551333215 date "2001-01-01" @default.
- W1551333215 modified "2023-09-27" @default.
- W1551333215 title "[9] Rabphilin-3: A target molecule for Rab3 small G proteins" @default.
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- W1551333215 doi "https://doi.org/10.1016/s0076-6879(01)29068-5" @default.
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