In Silico and In Vitro Studies on the Protein-Protein Interactions between Brugia malayi Immunomodulatory Protein Calreticulin and Human C1q
Figure 8
BmCRT binds at the globular head region of C1q. (A) IgG inhibits binding of C1q to immobilized BmCRT.
Microtiter plate was coated with BmCRT(1 µg/ml) in corbonate buffer (pH 9.6). C1q (1 µg/ml) was preincubated with 0.25 to 5 times excess (w∶w) IgG for 2 h at room temperature before addition to wells and and incubate overnight at 4°C. Binding of BmCRT with C1q in presence of IgG was observed by taking OD at 490 nm as mentioned in method section. (B) SAP shows no effect on BmCRT-C1q binding. Preincubated C1q (1 µg/ml) with 20 to 400 times execces (w∶w) SAP for 2 h was added in BmCRT (1 µg/ml) coated wells and binding of BmCRT with C1q was observed at 490 nm. BSA taking in both assays as a control, show no effect on BmCRT-C1q complex formation. The results were analyzed by two-way ANOVA (***P<0.001). Assay was performed in triplicates. Bar represent the standard deviations of the mean.