Figure 1.

Lpg2603 is an active kinase in the presence of IP6. A, sequence logo built using 74 homologs of Lpg2603 depicting conserved motifs present in kinases including the Gly-rich loop and active site residues (numbering in parentheses corresponds to PKA). The logo for canonical kinases was built using 3998 homologs (Pfam domain PF00069). B, autoradiograph depicting the incorporation of ³²P from [γ-³²P]ATP into MBP by recombinant Lpg2603 WT or the inactive mutant, D201A, in the presence of different cofactors: IP6, PI4P, ubiquitin (Ub), calmodulin (CaM), globular actin, or 14-3-3 protein. Note that CaM migrates at the same molecular weight as MBP. The reactions were terminated after 10 min by the addition of EDTA, and the products were resolved by SDS-PAGE and visualized by Coomassie Blue staining (lower panel) and autoradiography (upper panel). The results are representative of three independent experiments.