Figure 1 | Scientific Reports

Figure 1

From: Effects of altered N-glycan structures of Cryptococcus neoformans mannoproteins, MP98 (Cda2) and MP84 (Cda3), on interaction with host cells

Figure 1

Secretory expression of MP98 and MP84 in C. neoformans WT and alg3Δ mutant strains. (A) Scheme of His-tagged MP98 and MP84 without GPI anchorage. Details about the ORF, promoter, and terminator sequences of the genes encoding MP98 and MP84 were obtained from Broad Institute (https://www.broadinstitute.org). Pfam and SMART were used for domain prediction. Signal sequence and GPI anchor sites were predicted using SignalP 4.1 Server and PredGPI. N- and O-linked glycosylation sites were predicted on the basis of NetNGlyc 1.0 Server, NetOGlyc 4.0 Server, and YinOYang 1.2 tool analysis. The native MP84 promoter was exchanged with the histone 3 (H3) promoter (PH3) to direct a high MP84 expression. (B) Coomassie Blue staining of the purified MPs from C. neoformans WT and alg3Δ strains. MPs were purified from cell culture supernatants and analyzed through SDS-PAGE with bovine serum albumin (1 μg BSA) for comparison. The raw data of Coomassie Blue staining in (B) was presented in Supplementary Information as Fig. S5. (C) HPLC profiles of neutral and acidic (*) N-glycans assembled on MP98 and MP84. M8, M5, and M7 represent Man8GlcNAc2, Man5GlcNAc2, and Man7GlcNAc2, respectively.

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