Skeletal Muscle Lysosomal Function via Cathepsin Activity Measurement
Muscle wasting or cachexia is commonly associated with aging and many diseases such as cancer, infection, autoimmune disorders, and trauma. Decrease in muscle mass, or muscle atrophy, is often caused by dysfunction of protein proteolytic systems, such as lysosomes, which regulate protein turnover and homeostasis. Lysosomes contain many hydrolases and proteases and, thus, represent the major organelle that control protein turnover. Recently, lysosomes have emerged as a signaling hub to integrate cellular functions of nutrient sensing and metabolism, autophagy, phagocytosis, and endocytosis, which are all related to tissue homeostasis. In this chapter, we describe the protocol used to measure lysosomal proteinase (cathepsins) activity in the skeletal muscle. A better understanding of lysosomal function in muscle homeostasis is critical in developing new therapeutic approaches to prevent muscle wasting.
KeywordsAtrophy Autophagy Enzyme kinetics Fluorimeter Muscle acid lysates (MAL) Protein degradation Skeletal muscle function
This work was supported by NIH grant R01GM092759 (to M.X.Z) and an Ohio State University intramural Lockwood Research grant (to P.H.L).