Rabphilin-3A binds to a Mr 115,000 polypeptide in a phosphatidylserine- and Ca2+-dependent manner

Mutsuo Miyazaki, Kozo Kaibuchi, Hiromichi Shirataki, Hideshi Kohno, Tomomi Ueyama, Junsuke Nishikawa, Yoshimi Takai

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17 Citations (Scopus)


Rabphilin-3A is a putative target protein for Rab3A/Smg 25A, which is a member of the Ras-related small GTP-binding protein and implicated in neurotransmitter release from the synapse. Rabphilin-3A is composed of two functionally different domains: the N-terminal Rab3A-binding and the C-terminal phosphatidylserine- and Ca2+-binding domains. The C-terminal domain has two copies of an internal repeat that are homologous to the C2 domains of protein kinase C, synaptotagmin, and phospholipase A2 and C-γl, which are known to bind phosphatidylserine and Ca2+. In this study, we attempted to identify the Rabphilin-3A-interacting molecule in bovine brain by use of an overlay assay technique. The 32P-labeled C-terminal fragment of Rabphilin-3A (281-704 amino acids) bound to a protein molecule with a Mr of about 115 kDa which was immobilized on a nitrocellulose sheet. This protein was highly purified and characterized. The binding of the 32P-labeled C-terminal fragment to this protein was dependent on both phosphatidylserine and Ca2+, and inhibited by an excess amount of the C-terminal fragment and the C2 domain fragment (396-704 amino acids) but not by the N-terminal fragment (1-280 amino acids). These results indicate that Rabphilin-3A binds to a protein molecule with a Mr of 115 kDa through the C2 domain in the presence of phosphatidylserine and Ca2+.

Original languageEnglish
Pages (from-to)29-36
Number of pages8
JournalMolecular Brain Research
Issue number1
Publication statusPublished - 01-1995
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cellular and Molecular Neuroscience


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