論文 - 詳細
| RRC ID | 80726 |
|---|---|
| 著者 | Hara Y, Ichiraku A, Matsuda T, Sakane A, Sasaki T, Nagai T, Horikawa K. |
| タイトル | High-affinity tuning of single fluorescent protein-type indicators by flexible linker length optimization in topology mutant. |
| ジャーナル | Commun Biol |
| Abstract |
Genetically encoded Ca2+ indicators (GECIs) are versatile for live imaging of cellular activities. Besides the brightness and dynamic range of signal change of GECIs, Ca2+ affinity is another critical parameter for successful Ca2+ imaging, as the concentration range of Ca2+ dynamics differs from low nanomolar to sub-millimolar depending on the celltype and organism. However, ultrahigh-affinity GECIs, particularly the single fluorescent protein (1FP)-type, are lacking. Here, we report a simple strategy that increases Ca2+ affinity through the linker length optimization in topology mutants of existing 1FP-type GECIs. The resulting ultrahigh-affinity GECIs, CaMPARI-nano, BGECO-nano, and RCaMP-nano (Kd = 17-25 nM), enable unique biological applications, including the detection of low nanomolar Ca2+ dynamics, highlighting active signaling cells, and multi-functional imaging with other second messengers. The linker length optimization in topology mutants could be applied to other 1FP-type indicators of glutamate and potassium, rendering it a widely applicable technique for modulating indicator affinity. |
| 巻・号 | 7(1) |
| ページ | 705 |
| 公開日 | 2024-6-8 |
| DOI | 10.1038/s42003-024-06394-0 |
| PII | 10.1038/s42003-024-06394-0 |
| PMID | 38851844 |
| PMC | PMC11162441 |
| MeSH | Calcium* / metabolism HEK293 Cells Humans Luminescent Proteins* / chemistry Luminescent Proteins* / genetics Luminescent Proteins* / metabolism Mutation* |
| IF | 4.165 |
| 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 | 69 |
| 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 | HeLa(RCB0007) 293(RCB1637) |