The G2019S pathogenic mutation disrupts sensitivity of leucine-rich repeat kinase 2 to manganese kinase inhibition

P>Mutations in leucine-rich repeat kinase-2 (LRRK2) are the most common cause of late-onset Parkinson disease. Previously, we showed that the G2019S pathogenic mutation can cause a dramatic increase (approximately 10-fold) in kinase activity, far above other published studies. A notable experimental difference was the use of Mn-ATP as a substrate. Therefore, the effects of metal cation-ATP cofactors on LRRK2 kinase activity were investigated. It is shown, using several divalent metal cations, that only Mg2+ or Mn2+ can support LRRK2 kinase activity. However, for wild-type, I2020T, and R1441C LRRK2, Mn2+ was significantly less effective at supporting kinase activity. In sharp contrast, both Mn2+ and Mg2+ were effective at supporting the activity of G2019S LRRK2. These divergent effects associated with divalent cation usage and the G2019S mutation were predominantly because of differences in catalytic rates. However, LRRK2 was shown to have much lower (approximately 40-fold) ATP K-m for Mn-ATP compared with Mg-ATP. Consequently, sub-stoichiometric concentrations of Mn2+ can act to inhibit the kinase activity of wild-type, but not G2019S LRRK2 in the presence of Mg2+. From these findings, a new model is proposed for a possible function of LRRK2 and the consequence of the G2019S LRRK2 pathogenic mutation.