The lectin-specific activity of Toxoplasma gondii microneme proteins 1 and 4 binds Toll-like receptor 2 and 4 N-glycans to regulate innate immune priming
Fig 1
Lectin activity of rMIC1 and rMIC4.
(A) Glycoarray of the native MIC1/MIC4 subcomplex (Lac+) and of the recombinant forms of MIC1 and MIC4. In total, 320 oligosaccharide probes were analysed by reading their fluorescence intensities, and the 20 best recognized glycans are shown. The results were represented as previously reported [18]. (B) The activity and inhibition assays were performed in microplates coated with glycoproteins with or without sialic acid, fetuin (black bars), or asialofetuin (white bars), separately. After coating, wild type or mutated rMIC1 and rMIC4, pre-incubated with PBS or their corresponding sugars, were added to the wells. Later, bound proteins were detected through the addition of serum from T. gondii-infected mice. Data in (B) are expressed as mean ±S.D. of triplicate wells and significance was calculated with ANOVA followed by Bonferroni's multiple comparisons test. *p<0.05. Data are representative of two (B) independent experiments. Gal: galactose; GalNAc: N-acetylgalactosamine; Glc: glucose; Man: mannose; Fuc: fucose; Neu5Ac: N-acetylneuraminic acid; wt: wild type protein; mut: protein with a mutation in the carbohydrate-recognition domain (CRD); ns: not significant.