Reference - Detail
| RRC ID | 50105 |
|---|---|
| Author | Katayama H, Kogure T, Mizushima N, Yoshimori T, Miyawaki A. |
| Title | A sensitive and quantitative technique for detecting autophagic events based on lysosomal delivery. |
| Journal | Chem Biol |
| Abstract |
We sought to develop a sensitive and quantitative technique capable of monitoring the entire flux of autophagy involving fusion of lysosomal membranes. We observed the accumulation inside lysosomal compartments of Keima, a coral-derived acid-stable fluorescent protein that emits different-colored signals at acidic and neutral pHs. The cumulative fluorescent readout can be used to quantify autophagy at a single time point. Remarkably, the technique led us to characterize an autophagy pathway in Atg5-deficient cells, in which conventional LC3-based autophagosome probes are ineffective. Due to the large Stokes shift of Keima, this autophagy probe can be visualized in conjunction with other green-emitting fluorophores. We examined mitophagy as a selective autophagic process; time-lapse imaging of mitochondria-targeted Keima and GFP-Parkin allowed us to observe simultaneously Parkin recruitment to and autophagic degradation of mitochondria after membrane depolarization. |
| Volume | 18(8) |
| Pages | 1042-52 |
| Published | 2011-8-26 |
| DOI | 10.1016/j.chembiol.2011.05.013 |
| PII | S1074-5521(11)00204-3 |
| PMID | 21867919 |
| MeSH | Animals Anthozoa / chemistry Anthozoa / genetics Autophagy* Autophagy-Related Protein 5 Cell Line Gene Deletion Green Fluorescent Proteins / analysis Green Fluorescent Proteins / genetics Luminescent Proteins / analysis* Luminescent Proteins / genetics Lysosomes / metabolism* Mice Microscopy, Confocal / methods* Microtubule-Associated Proteins / genetics Mitochondria / metabolism Mutagenesis, Site-Directed Recombinant Fusion Proteins / analysis Recombinant Fusion Proteins / genetics |
| IF | 5.915 |
| Times Cited | 237 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 21 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| DNA material | mKeima (h)/pCS2 (RDB15236) dKeima (h)/pCS2 (RDB15237) mt-mKeima (h)/pIND(SP1) (RDB15238) mt-mKeima/pcDNA3 (RDB15456). |