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Identification of Key Chaperone-Mediated Autophagy Molecules

Background of Key Chaperone-Mediated Autophagy Molecules

Chaperone-mediated autophagy (CMA) is a highly regulated cellular process that maintains cellular homeostasis through lysosome-mediated degradation of damaged proteins and organelles .Increased CMA activity is directly correlated with the amount of lysosome-associated membrane protein type 2A (LAMP-2A) protein present in the lysosomal membrane. LAMP-2A is a transmembrane protein component for protein translocation to the lysosome. The most studied LAMP-2A was found to be present in CMA complexes extracted from different tissues during CMA activation . In addition, its chaperone and co-chaperone complexes are involved with LAMP-2A in mediating the translocation of CMA substrates across the lysosomal membrane. Isolation of these components by their specific antibodies inhibits CMA activity. Therefore, it is necessary to address some of these salient questions in molecular detail to understand the complex and important cellular process of CMA.

Consequences of downregulated CMA activity. Fig. 1. Consequences of downregulated CMA activity. (Alfaro I E, et al., 2019)

Key Chaperone-Mediated Autophagy Molecules Identification Services

LAMP-2A is a single-span transmembrane protein with a highly glycosylated tubular lumen structural domain and a short cytoplasmic tail in which CMA substrates bind prior to translocation into the lumen. At CD BioSciences, we are committed to providing comprehensive analysis of LAMP-2A dynamics services to our clients worldwide. We offer NMR-based techniques to analyze LAMP-2A transmembrane(TM) structural domains and characterize LAMP-2A interactions with molecular chaperones and CMA substrates in micelles.

Our scientists develop customized processes for analyzing the structure and dynamics of the LAMP-2A transmembrane structural domain.

(1) Design of LAMP-2A constructs containing the TM structural domain and C-terminal cytoplasmic tail.

(2) Nuclear magnetic resonance spectroscopy analyzes the structure of the LAMP-2A transmembrane structural domain and monitors LAMP-2A and partner Hsc70 protein interactions.

(3) The effect of micelles on the buoyant molecular mass of TM-LAMP-2A micelle complexes was eliminated by ultracentrifugation.

(4) Chemical cross-linking experiments were performed in reconstituted TM-LAMP-2A in micelles to determine the quaternary structure and stoichiometry of the subunits of the homologous oligomeric protein complex.

(5) Nuclear magnetic resonance structure calculations.

Why Choose Us

  • Nuclear magnetic resonance techniques provide reliable data.
  • Mature transgenic mouse model platform.
  • Rapidly characterize the structure and dynamics of LAMP-2A transmembrane structural domains.
  • Fully customized analytical services for LAMP-2A kinetic analysis.
  • Our dedicated team has unique insights into chaperone-mediated autophagy.

CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.

Reference

  1. Alfaro I E, et al. (2019) Chaperone mediated autophagy in the crosstalk of neurodegenerative diseases and metabolic disorders[J]. Frontiers in Endocrinology. 9: 778.

For research use only, not intended for any clinical use.

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