1. Academic Validation
  2. Two Methods to Analyze LRRK2 Functions Under Lysosomal Stress: The Measurements of Cathepsin Release and Lysosomal Enlargement

Two Methods to Analyze LRRK2 Functions Under Lysosomal Stress: The Measurements of Cathepsin Release and Lysosomal Enlargement

  • Methods Mol Biol. 2021;2322:63-72. doi: 10.1007/978-1-0716-1495-2_7.
Maria Sakurai 1 Tomoki Kuwahara 2
Affiliations

Affiliations

  • 1 Department of Neuropathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
  • 2 Department of Neuropathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. [email protected].
Abstract

Leucine-rich repeat kinase 2 (LRRK2) is a causative gene product of autosomal-dominant Parkinson's disease and has been shown to play a role in lysosomal regulation. We have previously shown that endogenous LRRK2 recruited its substrates Rab8a and Rab10 onto overloaded lysosomes depending on their phosphorylation, which functioned in the suppression of lysosomal enlargement as well as the promotion of the exocytic release of lysosomal cathepsins. In this chapter, we introduce two methods to analyze cellular functions of LRRK2 upon exposure to lysosomal overload stress in RAW264.7 cells.

Keywords

Cathepsin; LRRK2; Lysosomes; Lysosomotropic agent; Rab.

Figures
Products