Mutations in the
Parkinson's disease (PD)-associated
protein leucine-rich repeat kinase 2 (LRRK2) commonly lead to a reduction of
GTPase activity and increase in
kinase activity. Therefore, strategies for drug development have mainly been focusing on the design of LRRK2
kinase inhibitors. We recently showed that the central RocCOR domains (Roc: Ras of complex
proteins; COR: C-terminal of Roc) of a bacterial LRRK2 homolog cycle between a dimeric and monomeric form concomitant with
GTP binding and hydrolysis. PD-associated mutations can slow down
GTP hydrolysis by stabilizing the
protein in its dimeric form. Here, we report the identification of two
Nanobodies (NbRoco1 and NbRoco2) that bind the bacterial Roco
protein (CtRoco) in a conformation-specific way, with a preference for the
GTP-bound state. NbRoco1 considerably increases the
GTP turnover rate of CtRoco and reverts the decrease in
GTPase activity caused by a PD-analogous mutation. We show that NbRoco1 exerts its effect by allosterically interfering with the CtRoco dimer-monomer cycle through the destabilization of the dimeric form. Hence, we provide the first proof of principle that allosteric modulation of the RocCOR dimer-monomer cycle can alter its
GTPase activity, which might present a potential novel strategy to overcome the effect of LRRK2 PD mutations.