Development of novel sensors for autophagosome

来源 :The 7th International Symposium on Autophagy 2015(第七届自噬国际研讨会 | 被引量 : 0次 | 上传用户:shy1201107
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  Autophagy, referred to as macroautophagy, is intracellular degradation pathway that autophagosomic-lysosomal degradation of bulk cytoplasmic product and misfolded or aggregated proteins.Formation or degradation of autophagosome, a hallmark of autophagy, is critical for autophagic process.Therefore, monitoring of autophagosome is essential for autophagy research.However, so far, expression of GFP/RFP-LC3 is only way to detect autophagosome in living cells, which has the limitations due to overexpression of LC3.Here,we tried to develop new sensing markers for autophagosome by monitoring endogenous LC3 using LC3 binding peptides (LBPs).Interestingly, S(N30)-EGFP-LBP which is modified by adding a membrane-binding peptide mostly was co-localized to RFP-LC3-positive autophagosome in HEK293T cells and in mouse cortical neurons, while GFP-LBP itself is not co-localized to RFP-LC3 positive autophagosome.We are currently modifying the LC3-binding peptides for monitoring autophagosome efficiently in living cells.
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