Supplementary Figure 3: Description of secondary-structural elements of PaTox^G and important amino acid residues.

(a) Sequence of P. asymbiotica PaTox^G and assignment of secondary structural elements. Amino acids analyzed by site-directed mutagenesis are marked with an arrow (Supplementary Table 1). (b) Topology diagram of PaTox^G with secondary structural elements (colored as in Fig. 3). The catalytic domain features a typical GT-A fold with nucleotide binding fold (blue) and acceptor binding fold (red), which are also referred to as individual domains. The adjacent 3-helix bundle is indicated in yellow. The first 17 N-terminal residues (monomer A; 18 residues monomer B), two short regions (A: residues 2300-2306, B: 2302-2305; A: 2386-2393, B: 2384-2392) and the C-terminal residues (A: 2421-2448; B: 2420-2448) are not resolved in the structure and likely to be disordered and shown as dotted lines. (c) Cellular effects of PaTox^G-mutants with exchanged catalytic core residues. Micrographs of HeLa cells treated for 4 h with PaTox^G wild type and the indicated catalytic core mutants (each 11 nM) in combination with PA (0.5 μg/ml) as delivery system. With the mutants PaTox^G-D2260A and PaTox^G-R2263A no cellular effects were observed. PA alone showed no effect (control). Scale bar, 50 μm. The experiment was repeated four times.