Assay for Identification of LRRK2 Inhibitors

Abstract

A method for assessing the effect of a test compound on LRRK2 in a cell-based system, the method comprising the steps of a) assessing the effect of exposing the cell-based system comprising LRRK2 to the test compound on the phosphorylation state of Ser910 and/or Ser935 of the LRRK2; and/or b) assessing the effect of exposing the cell-based system comprising LRRK2 to the test compound on the binding of the LRRK2 to a 14-3-3 polypeptide. The method may comprise or further comprise the step of assessing the effect of exposing the cell-based system comprising LRRK2 to the test compound on the subcellular location of LRRK2. The method is considered to be useful in assessing the effect of putative LRRK2 inhibitors in cell based systems, including in vivo systems.

1 . An antibody that binds specifically to LRRK2 phosphorylated at Ser910; or an antibody that binds specifically to LRRK2 phosphorylated at Ser935; or an antibody that binds specifically to LRRK2 that is not phosphorylated at Ser910; or an antibody that binds specifically to LRRK2 that is not phosphorylated at Ser935. 
     
     
         2 . A kit of parts comprising two or more of:
 1) an antibody that binds specifically to LRRK2 phosphorylated at Ser910, or an antibody that binds specifically to LRRK2 that is not phosphorylated at Ser910;   2) an antibody that binds specifically to LRRK2 phosphorylated at Ser935, or an antibody that binds specifically to LRRK2 that is not phosphorylated at Ser935;   3) a 14-3-3 polypeptide, or an antibody that specifically binds to a 14-3-3 polypeptide; and   4) a fluorescently labeled LRRK2 polypeptide, or polynucleotide encoding a fluorescently labeled LRRK2.   
     
     
         3 . A purified preparation or kit of parts comprising an LRRK2 polypeptide or polynucleotide or antibody binding specifically to LRRK2; and a 14-3-3 polypeptide or polynucleotide or antibody binding specifically to a 14-3-3 polypeptide. 
     
     
         4 . A method of characterising an LRRK2 mutant, the method comprising the steps of:
 a) assessing the phosphorylation state of Ser910 and/or Ser935 of the LRRK2 mutant; and/or   b) assessing the ability of the LRRK2 mutant to bind a 14-3-3 polypeptide.   
     
     
         5 . The method of  claim 4 , wherein the LRRK2 mutant is found in a patient with Parkinson's disease. 
     
     
         6 . The method of  claim 4 , wherein the method comprises, or further comprises, the step of assessing the subcellular location of the LRRK2 mutant, when expressed in a cell-based system.