Glycophosphatidylinositol-anchored proteins in Paramecium tetraurelia: possible role in chemoresponse.
We have begun to characterize the glycophosphatidylinositol (GPI)-anchored proteins of the Paramecium tetraurelia cell body surface where receptors and binding sites for attractant stimuli are found. We demonstrate here (i) that inositol-specific exogenous phospholipase C (PLC) treatment of the cell body membranes (pellicles) removes proteins with GPI anchors, (ii) that, as in P. primaurelia, there is an endogenous lipase that responds differently to PLC inhibitors compared with its response to an exogenous PLC, (iii) that salt and ethanol treatment of cells removes GPI-anchored proteins from whole, intact cells, (iv) that Triton X-114 phase partitioning shows that many GPI-anchored proteins are cleaved from pellicles by the endogenous lipase and enter the aqueous phase, and (v) that integral membrane proteins are not among those cleaved with PLC or in the salt/ethanol wash. Antisera against the proteins removed by the salt/ethanol washing procedure include antibodies against large surface antigens, which we confirm in this species to be GPI-anchored, and against an array of proteins of smaller molecular mass. These antisera specifically block the chemoresponse to some stimuli, such as folate, which we suggest are signaled through GPI-anchored receptors. Responses to cyclic AMP, which we believe involve an integral membrane protein receptor, and to NH4Cl, which requires no receptor, are not affected by the antisera. Antiserum against a mammalian GPI-anchored folate-binding protein recognizes a single band among the GPI-anchored salt and ethanol wash proteins. The same antiserum specifically blocks the chemoresponse to folate.
Paquette
C A
CA
University of Vermont, Department of Biology, Burlington 05405, USA.
Rakochy
V
V
Bush
A
A
Van Houten
J L
JL
D000818:Animals; D002628:Chemoreceptor Cells / Q000187:drug effects / Q000502:physiology*; D000242:Cyclic AMP / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000431:Ethanol / Q000494:pharmacology; D005492:Folic Acid / Q000494:pharmacology; D017261:Glycosylphosphatidylinositols / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D007106:Immune Sera / Q000494:pharmacology; D008049:Lipase / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D017830:Octoxynol; D010247:Paramecium / Q000737:chemistry* / Q000502:physiology*; D011092:Polyethylene Glycols / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D012996:Solutions; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000494:pharmacology
D004791:Enzyme Inhibitors; D017261:Glycosylphosphatidylinositols; D007106:Immune Sera; D008565:Membrane Proteins; D012996:Solutions; D000431:Ethanol; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D017830:Octoxynol; C010615:Nonidet P-40; D005492:Folic Acid; D000242:Cyclic AMP; D008049:Lipase; D010738:Type C Phospholipases