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. 1995 Mar 15;306(Pt 3):643–650. doi: 10.1042/bj3060643

Lack of glycosyl-phosphatidylinositol anchoring leads to precursor retention by a unique mechanism in Dictyostelium discoideum.

P C Pauly 1, C Klein 1
PMCID: PMC1136570  PMID: 7702555

Abstract

Gp80, a cell-adhesion molecule in Dictyostelium discoideum, is modified by N- and O-linked oligosaccharides, and a glycosylphosphatidylinositol (GPI) anchor. To identify sequences important for the addition of these modifications to gp80, we created a hybrid protein in which the C-terminal 136 amino acids of yeast invertase were replaced by the C-terminal 110 amino acids of gp80. When expressed in D. discoideum, this protein (Inv-gp80) was not GPI-anchored and was retained in a pre-Golgi compartment. Inv-gp80 did, however, display characteristics of a transmembrane protein, suggesting a novel mechanism for its retention. We also expressed a truncated version of the hybrid protein in which the C-terminal 22 amino acids of the Inv-gp80 were deleted. The truncated protein (Inv-gp80stop) was O-glycosylated and secreted. These observations indicate that the hybrid protein is not abnormally folded and demonstrate the importance of the C-terminal 22 amino acids in the retention of Inv-gp80. Together, the data suggest that oligomerization of the protein blocks its GPI anchoring.

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