論文 - 詳細
| RRC ID | 82131 |
|---|---|
| 著者 | Ohta Y, Furuta T, Nagai T, Horikawa K. |
| タイトル | Red fluorescent cAMP indicator with increased affinity and expanded dynamic range. |
| ジャーナル | Sci Rep |
| Abstract |
cAMP is one of the most important second messengers in biological processes. Cellular dynamics of cAMP have been investigated using a series of fluorescent indicators; however, their sensitivity was sub-optimal for detecting cAMP dynamics at a low concentration range, due to a low ligand affinity and/or poor dynamic range. Seeking an indicator with improved detection sensitivity, we performed insertion screening of circularly permuted mApple, a red fluorescent protein, into the cAMP-binding motif of PKA regulatory subunit Iα and developed an improved cAMP indicator named R-FlincA (Red Fluorescent indicator for cAMP). Its increased affinity (Kd = 0.3 μM) and expanded dynamic range (860% at pH 7.2) allowed the detection of subtle changes in the cellular cAMP dynamics at sub-μM concentrations, which could not be easily observed with existing indicators. Increased detection sensitivity also strengthened the advantages of using R-FlincA as a red fluorescent indicator, as it permits a series of applications, including multi-channel/function imaging of multiple second messengers and combinatorial imaging with photo-manipulation. These results strongly suggest that R-FlincA is a promising tool that accelerates cAMP research by revealing unobserved cAMP dynamics at a low concentration range. |
| 巻・号 | 8(1) |
| ページ | 1866 |
| 公開日 | 2018-1-30 |
| DOI | 10.1038/s41598-018-20251-1 |
| PII | 10.1038/s41598-018-20251-1 |
| PMID | 29382930 |
| PMC | PMC5789972 |
| MeSH | Calcium / metabolism Cells, Cultured Cyclic AMP / metabolism* Dictyostelium / metabolism* Fluorescent Dyes / chemistry* Humans Insulin-Secreting Cells / metabolism* Luminescent Proteins / metabolism* Molecular Imaging / methods* Red Fluorescent Protein Spectrometry, Fluorescence |
| IF | 3.998 |
| 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 | X(Twitter) |
| 各媒体での言及数の合計 | 17 |
| 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 | |
| ヒト・動物細胞 | 293T(RCB2202) |