Reference - Detail
| RRC ID | 48869 |
|---|---|
| Author | Sato M, Mori Y, Matsui T, Aoki R, Oya M, Yanagihara Y, Fukuda M, Tsuboi T. |
| Title | Role of the polybasic sequence in the Doc2alpha C2B domain in dense-core vesicle exocytosis in PC12 cells. |
| Journal | J Neurochem |
| Abstract |
The double C2 (Doc2) family is characterized by an N-terminal Munc13-1-interacting domain and C-terminal tandem C2 domains, and it comprises three isoforms, Doc2alpha, Doc2beta, and Doc2gamma, in humans and mice. Doc2alpha, the best-characterized, brain-specific isoform, exhibits Ca(2+)-dependent phospholipid-binding activity through its C2A domain, and the Ca(2+)-binding activity is thought to be important for the regulation of Ca(2+)-dependent exocytosis. In contrast to the C2A domain, however, nothing is known about the physiological functions of the C2B domain in regulated exocytosis. In this study, we demonstrated by a mutation analysis that the polybasic sequence in the C2B domain of Doc2alpha (306 KKSKHKTCVKKK 317) is required for binding of syntaxin-1a/synaptosome-associated protein of 25 kDa (SNAP-25) heterodimer. We also investigated the effect of Lys-to-Gln (named KQ) mutations in the polybasic sequence of the C2B domain on vesicle dynamics by total internal reflection fluorescence microscopy in PC12 cells. A Doc2alpha(KQ) mutant, which lacks binding activity toward syntaxin-1a/SNAP-25 heterodimer, significantly decreased the number of plasma membrane-docked vesicles before stimulation and strongly inhibited high-KCl-induced exocytosis from the plasma membrane-docked vesicles. These results indicate that the polybasic sequence in the C2B domain functions as a binding site for syntaxin-1a/SNAP-25 heterodimer and controls the number of 'readily releasable' vesicles in neuroendocrine cells. |
| Volume | 114(1) |
| Pages | 171-81 |
| Published | 2010-7-1 |
| DOI | 10.1111/j.1471-4159.2010.06739.x |
| PII | JNC6739 |
| PMID | 20403080 |
| MeSH | Amino Acid Sequence Animals Calcium-Binding Proteins / genetics Calcium-Binding Proteins / physiology* Cell Membrane / metabolism Cytosol / metabolism Exocytosis* Mice Molecular Sequence Data Mutation Nerve Tissue Proteins / genetics Nerve Tissue Proteins / physiology* PC12 Cells Protein Binding Protein Multimerization Protein Structure, Tertiary Protein Transport Rats Secretory Vesicles / metabolism* Synaptosomal-Associated Protein 25 / genetics Syntaxin 1 / genetics |
| IF | 4.066 |
| Times Cited | 17 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY NEUROSCIENCES |
| Altmetric score |
オルトメトリクス指標項目
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| Total number of mentions | 0 |
| Resource | |
| DNA material | pEF-T7-mouse Doc2 alpha (RDB15100) |