Biogenesis of gp80, the GPI-anchored cohesion protein in D. discoideum

We have begun to assess the contributions made by co- and post-translational modifications to the physical properties of the D. discoideum protein gp80, and its sensitivity to hydrolysis by an endogenous GPI-PLC like activity. Two-dimensional gel electrophoresis indicates that the presence of various gp80 isoforms reflects the differential processing of its N-linked oligosaccharides. The presence or absence of this modification does not, however, appear to alter the sensitivity of the protein to the endogenous GPI-PIPLC activity. gp80 synthesized in cells defective in O-glycosylation is found predominantly in the medium as proteolytic products. The relationship of the O-linked modifications to the endogenous GPI-PIPLC activity is not yet clear. However, their absence does not alter the sensitivity of gp80 to exogenous GPI-PIPLC. We attempted to inhibit the GPI anchoring of gp80 using mannoseamine. Although cells displayed a reversible block in the onset of aggregation and gp80 synthesis, this did not appear to reflect changes in anchoring. The contribution is now being examined in chimeric proteins.