Fig. 7.

Interaction of ghA mutants with gC1qR.

Microtiter wells were coated with 1.0, 0.5 or 0.25 μg/well of gC1qR in carbonate buffer and the plate incubated at 4 °C overnight. The next morning, contents were discarded and wells were blocked for 2 h with 2% BSA at 37 °C. After washing with PBS + 0.05% Tween, 2.5 μg/well of ghA wild type, R162A, R162E and MBP was added and the plate was incubated 1.5 h, 37 °C and 1.5 h at 4 °C. Wells then were washed and anti-MBP (1 g/ml) was added, incubated for 1 h 37 °C and the bound protein was detected using horseradish peroxidase conjugated IgG. After addition of OPD buffer, the color developed was read at 450 nm. Each data bar is a mean ± SD run in duplicates (n = 3).