論文 - 詳細
| RRC ID | 42849 |
|---|---|
| 著者 | Shibata H, Suzuki H, Yoshida H, Maki M. |
| タイトル | ALG-2 directly binds Sec31A and localizes at endoplasmic reticulum exit sites in a Ca2+-dependent manner. |
| ジャーナル | Biochem Biophys Res Commun |
| Abstract |
Intracellular localization of the penta-EF-hand Ca2+-binding protein ALG-2 in HeLa cells was investigated by immunofluorescent confocal microscopy using a polyclonal antibody. In addition to its presence in the nucleus, ALG-2 was found to be distributed in a punctate pattern in the cytoplasm, where it was partly co-stained with an endoplasmic reticulum (ER) exit site marker p125. In vitro GST pull down analysis demonstrated that ALG-2 and its alternatively spliced isoform interact with the COPII component Sec31A in a Ca2+-dependent manner, and a biotin-labeled ALG-2 overlay assay revealed direct binding of ALG-2 to Sec31A. Biochemical and immunofluorescent microscopic analyses showed that ALG-2 was enriched at the Sec31A-localizing membrane compartments upon stimulation with the Ca2+ ionophore A23187. In contrast, treatment of cells with the membrane-permeant Ca2+ chelator BAPTA-AM led to a dispersion of ALG-2 throughout the cells and to a significant loss of Sec31A in the perinuclear region. These findings establish Sec31A as a novel target for ALG-2 and provide a framework for studies on the roles of ALG-2 in ER-Golgi transport. |
| 巻・号 | 353(3) |
| ページ | 756-63 |
| 公開日 | 2007-2-16 |
| DOI | 10.1016/j.bbrc.2006.12.101 |
| PII | S0006-291X(06)02763-X |
| PMID | 17196169 |
| MeSH | Animals Apoptosis Regulatory Proteins / immunology Apoptosis Regulatory Proteins / metabolism* Calcimycin / pharmacology Calcium / physiology* Calcium-Binding Proteins / immunology Calcium-Binding Proteins / metabolism* Carrier Proteins / metabolism* EF Hand Motifs Egtazic Acid / analogs & derivatives Egtazic Acid / pharmacology Endoplasmic Reticulum / drug effects Endoplasmic Reticulum / metabolism* Endoplasmic Reticulum / ultrastructure HeLa Cells Humans Microscopy, Confocal Microscopy, Fluorescence Rabbits Vesicular Transport Proteins |
| IF | 2.985 |
| 引用数 | 54 |
| WOS 分野 | BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| リソース情報 | |
| ヒト・動物細胞 | HeLa(RCB0007) |