Recent studies have revealed how AMPK is activated inside the cell and animal tissues: in response to low glucose, AXIN tethers LKB1, by virtue of their constitutive association, to AMPK located on the surface of late endosome/lysosome. Importantly, the lysosomal v-ATPase (vacuolar ATPase)-Ragulator complex, when primed by glucose starvation or concanamycin A, facilitates AXIN/LKB1 to interact with AMPK. Here, we describe the experimental procedures of the assays for detecting the translocation of AXIN/LKB1 or the assembly of the AXIN-based AMPK-activating complexes on the late endosome/lysosome. The methods in this chapter will be useful for determining whether various metabolic stresses or pharmacological stimuli activate AMPK via the v-ATPase-Ragulator-AXIN/LKB1 axis, which also concomitantly inactivates mTORC1. Detailed protocols for determining the levels of adenylates are also described.
CITATION STYLE
Zhang, C. S., Li, M., Zong, Y., & Lin, S. C. (2018). Determining AMPK activation via the lysosomal v-ATPase-ragulator-AXIN/LKB1 axis. In Methods in Molecular Biology (Vol. 1732, pp. 393–411). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7598-3_25
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