Fig. 1.

Dual channel image of total protein amount (Flamingo fluorescent dye-stained; green) and phosphorylated proteins (Pro-Q Diamond-stained; red) of stationary phase M. pneumoniae cells. About 125 μg of a total protein extract of M. pneumoniae grown in modified Hayflick medium supplemented with glucose were subjected to two-dimensional gel electrophoresis using either an 18-cm IPG strip with a linear pH gradient of pI 6–11 (A) or pI 4–7 (B) in the first dimension. Dual channel images were obtained using the software DECODON Delta2D 3.6. Spots with high Pro-Q Diamond/Flamingo log ratios representing phosphorylated proteins (9) are indicated on the gel. Multiple protein spots corresponding to the same original protein are distinguished by appended digits (e.g. Tuf-1 and Tuf-2). Protein spots were cut from the gel and identified by MS/MS (see Table I and supplemental Table S3).