Background of Lysosomal Activation During Autophagy
Macroautophagy is a cellular catabolic process in response to starvation or other stress conditions, whereby cellular components are phagocytosed into autophagosomes and ultimately delivered to lysosomes for degradation. Lysosomes play a key role in performing autophagy as a degrading acidic organelle. Autolysosomes generated by the fusion of smaller acidic lysosomes with larger autophagosomes require some form of functional adjustment to maintain an optimal acidic pH to perform hydrolytic functions to digest luminal contents. Currently, the biogenesis of autophagosomes in the early stages of autophagy is relatively well studied, and the functional changes of lysosomes during autophagy remain largely unknown. Understanding the activation of lysosomal function during autophagy and determining the role of mTORC1 in this process is of great biological and medical importance.
Fig. 1. Regulation of lysosomes in the course of autophagy. (Shen H M, et al., 2014)
Lysosomal Activation During Autophagy Analysis Services
Studies have shown that the CLEAR (Coordinated Lysosomal Expression and Regulation) gene network and the transcription factor EB (TFEB) are thought to be major regulators of lysosomal biogenesis, function and autophagy, controlling gene expression through transcription. Starvation-induced inactivation of mTORC1 removes its inhibition of TFEB and autophagic flux is supported by elevated lysosomal activity. For many years, CD BioSciences has been committed to providing analytical services for lysosomal activation during autophagy to our global customers.
Our scientists have adopted a variety of strategies to explore the functional changes of lysosomes during autophagy.
(1) Analyze the role of mTORC1 as a negative regulator of lysosomal function.
(2) Analyze changes in mTOR activity and lysosomal localization in cells under starvation.
(3) Analysis of proteins involved in autophagosome-lysosome fusion, endosomal sorting complex (ESCRT), microtubules, Rab7 , soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) and lysosomal V-ATPase.
The technical platform we used to assess changes in lysosomal function in autophagy.
- Measurement of lysosomal acidification (using LysoTracker and LysoSensor).
- Cathepsin enzymatic activity (using Magic-Red reagent combined with flow cytometry).
- Lysosomal protein degradation ability (using DQ-BSA assay).
Why Choose Us
- Fully customizable services.
- Unique insights into autophagy-lysosome fusion from a team of professionals.
- Technical support for the control of lysosomal function in autophagy.
- Rapidly help you analyze the activation of lysosomal function during autophagy and determine the role of mTORC1 in this process.
- Assisting in the drug development of inhibitors of mTOR.
- Predicting possible molecular targets of mTORC1 at the lysosome.
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Reference
- Shen H M, Mizushima N. (2014) At the end of the autophagic road: an emerging understanding of lysosomal functions in autophagy[J]. Trends in biochemical sciences. 39(2): 61-71.