Methods to monitor chaperone-mediated autophagy.
Chaperone-mediated autophagy (CMA) is a selective type of autophagy responsible for the lysosomal degradation of soluble cytosolic proteins. In contrast to other forms of autophagy where cargo is sequestered and delivered to lysosomes through membrane fusion/excision, CMA substrates reach the lysosomal lumen after direct translocation across the lysosomal membrane. CMA is part of the cellular quality control systems and as such, essential for the cellular response to stress. CMA activity decreases with age, likely contributing to the accumulation of altered proteins characteristic in tissues from old organisms. Furthermore, impairment of CMA underlies the pathogenesis of certain human pathologies such as neurodegenerative disorders. These findings have drawn renewed attention to CMA and a growing interest in the measurement of changes in CMA activity under different physiological and pathological conditions. In this chapter we review the different experimental approaches used to assess CMA activity both in cells in culture and in different organs from animals.
Kaushik
Susmita
S
Department of Developmental and Molecular Biology, Marion Bessin Liver Research Center, Institute for Aging Research, Albert Einstein College of Medicine, Bronx, New York, USA.
Cuervo
Ana Maria
AM
D000818:Animals; D001343:Autophagy / Q000502:physiology*; D050883:HSC70 Heat-Shock Proteins / Q000378:metabolism; D015151:Immunoblotting; D052119:Lysosomal-Associated Membrane Protein 2 / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic
D050883:HSC70 Heat-Shock Proteins; D052119:Lysosomal-Associated Membrane Protein 2