Reactivity of fungal GIPCs with MEST-3. Fungal GIPCs were purified by a combination of chromatography in DEAE-Sephadex, silica gel 60, HPLC and preparative HPTLC. HPTLC was developed in solvent A. Panel A, stained with orcinol/H2SO4 and panel B, immunostaining with MEST-3. Lane 1, GIPC Pb-2 from mycelium form of P. brasiliensis; lane 2, acidic GSLs from mycelium form of P. brasiliensis; lane 3, acidic GSLs from yeast form of P. brasiliensis (Pb); lane 4, acidic GSLs from hyphae of A. fumigatus (Af); lane 5, acidic GSLs from hyphae of A. nidulans (An); lane 6, acidic GSLs from mycelium form of H. capsulatum (Hc); lane 7, acidic GSLs from yeast form of H. capsulatum; lane 8, acidic GSLs from mycelium form of S. schenckii (Sc); lane 9, acidic GSLs from yeast form of S. schenckii; lane 10, acidic GSLs from the edible mushroom Agaricus blazei (Ab). Arrows indicates chromatographic migration of Pb-2, Af-2, An-2, Hc-Y2 and Ss-Y2. Panel C, GIPCs (first well 0.1 μg) were serially double diluted in ethanol and adsorbed on a 96-well plate. MEST-3 (100 μl) was added and incubated overnight at 4°C. The amount of antibody bound to GSLs was determined by incubation with rabbit anti-mouse IgG (2 h) and 105 cpm of 125I-labeled protein A in 1% BSA. Pb-2 from yeast (closed square) and from mycelium (closed triangle) forms of P. brasiliensis; Ss-Y2 (open circle) from yeast form of S. schenckii; Af-2 (open triangle) from A. fumigatus, Hc-Y2 (open inverted triangle) from yeast forms of H. capsulatum, Pb-3 (closed inverted triangle) from yeast and Pb-3 (closed diamond) from mycelium forms of P. brasiliensis and Ss-M2 (open diamond) from mycelium forms of S. schenckii.
Toledo et al. BMC Microbiology 2010 10:47 doi:10.1186/1471-2180-10-47