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. Author manuscript; available in PMC: 2015 Feb 1.
Published in final edited form as: Environ Microbiol. 2013 Aug 19;16(2):359–381. doi: 10.1111/1462-2920.12223

Fig. 6.

Fig. 6

Legionella KaiC conserves autokinase activity but is not dephosphorylated by KaiB.

A. Western blot using an α-histidine antibody against the purified Legionella 6xHis-LpKaiC (65 kDa) and S. elongatus 6xHis-SeKaiC (58 kDa) proteins (lower panel). Autophosphorylation of SeKaiC and LpKaiC incubated with 5 μCi [γ-32P]-ATP for 1 h at 30°C (upper panel).

B. Kinetics of autophosphorylation and dephosphorylation of LpKaiC and SeKaiC in presence of KaiB or KaiB-T of L. pneumophila. Recombinant KaiC proteins were incubated with KaiB or KaiB-T, respectively and 5 μCi [γ -32P]-ATP at 30°C for 24 h. Samples were taken at 4 h, 8 h and 24 h of incubation and subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis analyses. The phosphorylation state was detected by autoradiography (upper panel) and Western blot of the reaction mixtures with α-His as loading control (lower panel).