Cleavage within Reelin repeat 3 regulates the duration and range of the signaling activity of Reelin protein
| Autor: | Atsushi Baba, Kazutomo Sasaki, Arisa Hisanaga, Mitsuharu Hattori, Mari Koie, Takao Kohno, Takana Kamei, Mengyan Deng, Kyoko Okumura |
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| Rok vydání: | 2014 |
| Předmět: |
Male
Proline Endosome Cell Adhesion Molecules Neuronal Blotting Western Molecular Sequence Data Nerve Tissue Proteins Endosomes Protein degradation Cleavage (embryo) Biochemistry CD49c Mice Neurobiology Extracellular Animals Humans Reelin Amino Acid Sequence Molecular Biology Cells Cultured Neurons Aspartic Acid Extracellular Matrix Proteins Binding Sites biology Sequence Homology Amino Acid Serine Endopeptidases Cell Biology DAB1 Molecular biology Cell biology Reelin Protein HEK293 Cells nervous system Mutation Proteolysis biology.protein Female Signal transduction Extracellular Space Peptide Hydrolases Signal Transduction |
| Zdroj: | The Journal of biological chemistry. 289(18) |
| ISSN: | 1083-351X |
| Popis: | Reelin is a secreted glycoprotein that plays essential roles in the brain. Reelin is specifically cleaved at two distinct sites, called N-t and C-t, with the former being the major one. N-t cleavage can occur both in the extracellular space and in the endosomes, although the physiological importance of endosomal N-t cleavage has not been investigated. In this study, we first determined the exact N-t cleavage site catalyzed by a protease secreted by cerebral cortical neurons. Cleavage occurred between Pro-1244 and Ala-1245 within Reelin repeat 3. A Reelin mutant in which Pro-1244 was replaced with aspartate (Reelin-PD) was resistant to a protease secreted by cultured cerebral cortical neurons, and its biological activity stayed active longer than that of wild-type Reelin. Interestingly, Reelin-PD remained in the intracellular compartments longer than wild-type Reelin and persistently activated downstream signaling. Therefore, N-t cleavage of Reelin is required for halting the signaling machinery in the extracellular space as well as within endosomes of target neurons. We established a monoclonal antibody specific to uncleaved Reelin protein and found that it is localized in the vicinity of Reelin-producing cells, whereas the N-terminal fragment diffuses, or is transported, to distant regions. These data demonstrate that N-t cleavage of Reelin plays critical roles in regulating the duration and range of Reelin functions both in the extracellular milieu and in the intracellular compartments. |
| Databáze: | OpenAIRE |
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