EDTA treatment alters protein glycosylation in the cellular slime mold Dictyostelium discoideum.
We have found that treatment of cells with EDTA resulted in the accumulation of lower molecular weight forms of two cell-type-specific glycoproteins. These new glycoproteins lacked a developmentally regulated glycoantigen defined by monoclonal antibody 54.2. Since EDTA dissociated the cells, the possible involvement of cell separation was tested by immobilizing cells in soft agarose. Glycoantigen expression on these proteins was found to be dependent on cAMP and high oxygen tension but not on cell contact, and was reversibly sensitive to EDTA regardless of the state of cell association. The EDTA effect was mimicked by other soluble, but not particulate, membrane impermeable chelators, could be competed by Zn2+ better than Mg2+, and appeared to involve an intracellular mechanism. Studies with [14C]EDTA showed that EDTA equilibrated with a cellular compartment in a temperature-dependent, Zn2+-insensitive fashion with half-time kinetics of loading and unloading of 30-40 min. If the compartment was assumed to be labeled with the same concentration of EDTA as was present extracellularly, calculations showed that its volume was circa 2% of the total cell volume. This compartment probably consists of intracellular vesicles based on the similar labeling of this compartment with a bulk fluid phase marker, inulin. The data suggest that this step in glycosylation, which was found to be delayed 1 or more hours subsequent to protein synthesis, involves an intracellular, transition metal ion-dependent process which can be modulated by chelators entering the cell through the endocytic pathway.
D002413:Cations, Divalent; D002448:Cell Adhesion; D002449:Cell Aggregation; D004023:Dictyostelium / Q000166:cytology / Q000502:physiology*; D004492:Edetic Acid / Q000494:pharmacology*; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D007158:Immunologic Techniques; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational / Q000187:drug effects*
D002413:Cations, Divalent; D006023:Glycoproteins; D004492:Edetic Acid