Fig. 2.

Reconstitution of PglA-mediated PilA glycosylation in E. coli. Immunoblotting of whole-cell lysates from strains expressing PilA and gonococcal oligosaccharyltransferase PglO (A) and F. tularensis subsp. tularensis strain SCHU S4 PglA (B). The undecaprenylphosphate-linked 2,4-diacetamido-2,4,6-trideoxyhexose (DATDH) precursor was expressed by virtue of the pglFDCD core locus from N. gonorrhoeae. Immunoblotting utilized polyclonal anti-PilA serum and the monoclonal antibody npg1 that recognizes an epitope imparted by the presence of the DATDH sugar. Plus and minus signs denote the proteins expressed (+) and not expressed (−) in the backgrounds tested.