Fig. 1.

PKM1 and PKM2 interact with laforin. a Yeast substrate-trapping system with LexA-laforin C266S was used to identify putative partners from a human brain library. Five clones corresponding to different lengths of PKM1 and PKM2 were selected (in brackets the amino acids included in the clones corresponding either to PKM1 of PKM2). A diagram of the different domains in PKMs is presented (black bar indicates the region that is different between PKM1 and PKM2). The strength of the interaction of these clones with either LexA-laforin C266S or LexA-laforin wild type was measured (β-galactosidase activity). b Co-immunoprecipitation analysis of PKM1 and PKM2 with laforin. HEK293 cells were transfected with the indicated plasmids. Crude extracts and co-immunoprecipitation analyses with anti-FLAG antibody was performed as indicated in Methods. Proteins in the immunoprecipitates and in the crude extracts (CE, 25 μg) were analyzed by western blotting using the indicated antibodies