論文 - 詳細
| RRC ID | 28665 |
|---|---|
| 著者 | Ohkura M, Matsuzaki M, Kasai H, Imoto K, Nakai J. |
| タイトル | Genetically encoded bright Ca2+ probe applicable for dynamic Ca2+ imaging of dendritic spines. |
| ジャーナル | Anal Chem |
| Abstract |
G-CaMP is a Ca2+ probe based on a single green fluorescent protein (GFP). G-CaMP shows a large fluorescence increase upon Ca2+ binding, but its fluorescence is dim and pH sensitive, similar to other single GFP-based probes. Here we report an improved G-CaMP, named G-CaMP1.6, which enables easier detection of intracellular Ca2+ signals. G-CaMP1.6 was approximately 40 times more fluorescent than G-CaMP, mainly due to an increase in quantum yield. Furthermore, compared with G-CaMP, G-CaMP1.6 had not only a lower pH sensitivity but also a higher selectivity for divalent cations having an ionic radius similar to Ca2+. Ca2+ sensitivity of G-CaMP1.6 (Kd = 146 nM, Hill coefficient = 3.8, Fmax/Fmin = 4.9) was slightly shifted toward higher affinity compared with that of G-CaMP. When expressed in mammalian cells, G-CaMP1.6 showed large fluorescence changes with drug applications. Notably, local Ca2+ changes in such tiny structures as dendritic spines of neurons were successfully observed with G-CaMP1.6, this being the first observation using a GFP-based probe. Additional mutations in Ca2+-binding sites of G-CaMP1.6 shifted the affinity for Ca2+ and reduced the Ca2+-buffering effect. G-CaMP1.6-CaM(E140K), which has a mutation in the Ca2+ binding site, is an improved probe with its increased brightness and reduced Ca2+-buffering capacity. |
| 巻・号 | 77(18) |
| ページ | 5861-9 |
| 公開日 | 2005-9-15 |
| DOI | 10.1021/ac0506837 |
| PMID | 16159115 |
| MeSH | Animals Calcium / analysis* Calcium / metabolism* Cell Line Dendritic Spines / genetics* Dendritic Spines / metabolism* EF Hand Motifs Genetic Variation / genetics Hippocampus / metabolism Humans Rats Sensitivity and Specificity Temperature |
| IF | 6.785 |
| 引用数 | 79 |
| WOS 分野 | CHEMISTRY, ANALYTICAL |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 遺伝子材料 | pN1-G-CaMP1.6 (RDB06749) |