Figure 5

Influence of ions and lysine analog ACA on binding of rMpn_Ef-Tu to plasminogen and degradation of human fibrinogen and vitronectin by activated plasminogen. (a) Microtitre plate wells were coated with rMpn_Ef-Tu and incubated with plasminogen and increasing concentrations of either NaCl (‘ionic interactions’) or ε-aminocaproic acid (‘ACA’). Bound plasminogen was detected with anti-plasminogen antibodies. In control experiments M. pneumoniae cells were coated onto microtitre plates and incubated with plasminogen and increasing concentrations of either NaCl or ACA. Bars represent standard deviation of eight replicates. (b) Fibrinogen or vitronectin was mixed with either urinary plasminogen activator (uPA) or tissue plasminogen activator (tPA) and added to microtitre plates previously coated with rMpn_Ef-Tu and plasminogen. Samples were separated by SDS-PAGE, blotted onto nitrocellulose membrane and probed with anti-vitronectin and anti-fibrinogen antisera. Lane 1 is at 0 hours, lane 2 is after over-night incubation without plasminogen, and lane 3 is after over-night incubation with plasminogen. Full length blots can be seen in Figure S9.