Malattia di Parkinson: ruolo della chinasi LRRK2 nella fosforilazione di Rab10

Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized by the selective loss of dopaminergic neurons in the Substantia Nigra pars compacta in the midbrain, resulting in a series of typical motor and non-motor symptoms. The etiology of PD remains unclear; most of PD cases (around 90%) are idiopathic, but mutations in at least 20 identified genes have been linked to monogenic form of the disease. The attention is here focused on LRRK2 (Leucine-Rich Repeat Kinase 2) gene, which encodes for a kinase protein that is responsible for the phosphorylation of a subset of Rab GTPases. LRRK2 through the phosphorylation of Rab proteins plays an important role in several cellular processes such as the regulation of vesicles trafficking, cytoskeleton dynamics, autophagy and lysosomal degradation. Recently it has also been discovered that LRRK2 is involved in the regulation of immune response. Mutations in lrrk2, resulting in an enhanced kinase activity, are the most common cause of monogenic PD and one of the most important risk factors for the idiopathic forms of the disease. The aim is to validate an endogenous system for assessing LRRK2 activity. Thanks to different techniques such as immunocytochemistry and western blot, changes in LRRK2 activity after its inhibition with MLi2 are evaluated by measuring the phosphorylation of Rab10, which is one of its best-characterized substrates.