FIGURE 2.
ZmpI-mediated release of the cell wall-associated CD2831 protein into the culture supernatant. A, schematic representation of CD2831. The relevant characteristics are a signal peptide (SP), two putative collagen-binding domains (CBD), a CnaB-like domain, and a C-terminal putative sorting signal comprising a PPKTG sorting motif, a hydrophobic domain (in gray), and a positively charged tail (+). AA, amino acids. B, supernatant (Sn), cell wall (CW), membrane (Mb), and cytosolic (Cy) compartment of wild type (WT), Ptet-srtB (overexpresses the sortase in the presence of ATc), and zmpI mutant (ΔzmpI) strains, carrying pJKP041 (expresses CD2831 in the presence of ATc) and cultivated in the presence of 100 ng/ml ATc (+ATc), were analyzed by Western blotting with anti-CD2831 polyclonal antibodies. Supernatant and whole cell lysate (WL) fractions of ΔzmpI pJKP041 strain cultivated in the absence of ATc (−ATc) were also analyzed as negative controls.