Detection of Substrate Phosphorylation of LRRK2 in Tissues and Cultured Cells

Methods Mol Biol. 2021:2322:53-61. doi: 10.1007/978-1-0716-1495-2_6.

Abstract

Recent studies revealed that leucine-rich repeat kinase 2 (LRRK2) phosphorylates several Rab proteins under physiological conditions. Mutations linked with familial Parkinson's disease cause an abnormal increase in the Rab phosphorylation, which has not been elucidated in an in vitro kinase assays where artificial peptide substrates are often used. Here, we provide protocols for detecting the LRRK2 activity in tissues and cultured cells using Rab phosphorylation as a readout.

Keywords: Leucine-rich repeat kinase 2; Parkinson disease; Phos-tag; Phosphorylation; Rab.

MeSH terms

  • 3T3 Cells
  • A549 Cells
  • Animals
  • Cell Line
  • Cell Line, Tumor
  • Humans
  • Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / genetics
  • Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / metabolism*
  • Mice
  • Mutation / genetics
  • Parkinson Disease / genetics
  • Parkinson Disease / metabolism
  • Phosphorylation / physiology*

Substances

  • LRRK2 protein, human
  • Leucine-Rich Repeat Serine-Threonine Protein Kinase-2