Extended Data Fig. 7. The mechanism of SidJ induced CaM dependent self-AMPlyation and SdeA glutamylation.

a. SidJ induces self-AMPylation in a CaM dependent manner. SidJ was incubated with ³²P-α-ATP, Mg²⁺, with or without CaM for 2 h at 37°C. After separation by SDS-PAGE, the incorporation of ³²P-α-ATP was detected by autoradiography.

b. SdeA glutamylation by SidJ interferes with SidJ self-AMPylation. SidJ was incubated with ³²P-α-ATP, Mg²⁺, CaM for 2 h at 37°C. When needed L-Glu, SdeA, SdeA_E860A were supplemented. After separation by SDS-PAGE, the incorporation of ³²P-α-ATP was detected by autoradiography.

c. SdeA glutamylation by SidJ accelerates ATP hydrolysis and AMP release. SidJ was incubated with indicated components for 2 h at 37°C. Samples were analyzed by HPLC. AMP and ATP were used as standard. In panels a-c, data shown were one representative from at least three independent experiments that had similar results.

d. Schematic model of SidJ induced glutamylation and AMPylation. SidJ incudes glutamylation on SdeA_E860 when ATP and L-Glu are supplemented in reaction. In reactions missing L-Glu or modifiable SdeA, SidJ induces self-AMPylation.