Fig. 3.

Only IIC and IID of the man-PTS are required for complex-formation with fLciA and lactococcin A. The presence (+) or absence (−) of ptnAB, ptnC, ptnD, and flciA in the various clones (Bact. clone) is indicated in the (Top) as is the presence (+) or absence (−) of added lactococcin A (50 BU/ml) and nisin (0.2 or 1 ng/ml for induction of the cloned genes). After overnight growth, fLciA and its copurified proteins were collected by immunoprecipitation and analyzed by SDS/PAGE and silver staining (Middle). For immunodetection of fLciA with the antibody M2 (α-fLciA; only area containing fLciA is shown here), purified samples were diluted 5- or 25-fold (∗; to avoid oversaturated signals) before 10 μl of each diluted sample was analyzed by Western blotting. (Bottom). Coprecipitated bacteriocin activity was analyzed by applying 5 μl of the eluted fractions directly onto lawns of the indicator stain Il1403. Growth inhibition is seen as clear zones. Filled arrowheads denote the man-PTS components, and open arrowheads (in bacteriocin-treated B268 and B607) indicate a protein that was coprecipitated only when the complete man operon was expressed.