Figure 8.
Hypoglycosylation of Gas1 stimulates shedding by native Yps1. A) Schematic view of Gas1p glycosylation profile and putative cleavage sites as depicted for Yps1 (Figure 7A). B) Cell extracts (C; 5.0 × 106 cells equivalent) and concentrated supernatants (S; 10-times more material compared to cell extracts) from 24 h cultures at pH 6.0 of the indicated yeast mutants (all strains are also yps1Δ) either transformed (pYPS1) or untransformed with plasmid Yps1 were separated by 10% SDS-PAGE and immunodetected with a Gas1 antiserum. Asterisks point to faint discrete Gas1 species detected above the major 80 kDa species in the extracellular milieu. The percentage of total Gas1-immunoreactive material (%T) associated with cell extracts and supernatants is indicated for all strains.