Reference - Detail
| RRC ID | 32583 |
|---|---|
| Author | Fukuda M. |
| Title | Slp4-a/granuphilin-a inhibits dense-core vesicle exocytosis through interaction with the GDP-bound form of Rab27A in PC12 cells. |
| Journal | J Biol Chem |
| Abstract |
Slp4-a (synaptotagmin-like protein 4-a)/granuphilin-a is specifically localized on dense-core vesicles in PC12 cells and negatively controls dense-core vesicle exocytosis through specific interaction with Rab27A via the N-terminal Slp homology domain (SHD) (Fukuda, M., Kanno, E., Saegusa, C., Ogata, Y., and Kuroda, T. S. (2002) J. Biol. Chem. 277, 39673-39678). However, the mechanism of the inhibition by Slp4-a has never been elucidated at the molecular level and is still a matter of controversy. In this study, I discovered an unexpected biochemical property of Slp4-a, that Slp4-a, but not other Rab27 effectors reported thus far, is capable of interacting with both Rab27A(T23N), a dominant negative form that mimics the GDP-bound form, and Rab27A(Q78L), a dominant active form that mimics the GTP-bound form, whereas Slp4-a specifically recognizes the GTP-bound form of Rab3A and Rab8A and does not recognize their GDP-bound form. I show by deletion and mutation analyses that the TGDWFY sequence in SHD2 is essential for Rab27A(T23N) binding, whereas SHD1 is involved in Rab27A(Q78L) binding. I further show by immunoprecipitation and cotransfection assays that Munc18-1, but not syntaxin IA, directly interacts with the C-terminal domain of Slp4-a in a Rab27A-independent manner. Expression of Slp4-a mutants that lack Rab27A(T23N) binding activity (i.e. specific binding to Rab27A(Q78L)) completely reverses the inhibitory effect of the wild-type Slp4-a on high KCl-dependent neuropeptide Y secretion in PC12 cells. The results strongly indicate that interaction of Slp4-a with the GDP-bound form of Rab27A, not with syntaxin IA or Munc18-1, is the primary reason that Slp4-a expression inhibits dense core vesicle exocytosis in PC12 cells. |
| Volume | 278(17) |
| Pages | 15390-6 |
| Published | 2003-4-25 |
| DOI | 10.1074/jbc.M213090200 |
| PII | S0021-9258(19)30349-7 |
| PMID | 12590134 |
| MeSH | Amino Acid Substitution Animals Base Sequence Carrier Proteins / metabolism Carrier Proteins / physiology* DNA Mutational Analysis Exocytosis* / genetics Guanosine Diphosphate / metabolism Mice Molecular Sequence Data Munc18 Proteins Nerve Tissue Proteins / metabolism Neuropeptide Y / metabolism PC12 Cells / metabolism* Protein Binding / genetics Protein Binding / physiology Protein Structure, Tertiary Rats Secretory Vesicles* Vesicular Transport Proteins / metabolism rab GTP-Binding Proteins / genetics rab GTP-Binding Proteins / metabolism* rab27 GTP-Binding Proteins |
| IF | 4.238 |
| Times Cited | 52 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Wikipedia |
| Total number of mentions | 5 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| DNA material | Slp4-a (RDB03421) pEF-T7-mouse Slac2-b (RDB15095) pEF-FLAG-mouse Munc18-1 (RDB15177). |