Cleavage with phospholipase of the lipid anchor in the cell adhesion molecule, csA, from Dictyostelium discoideum.
A cell adhesion molecule, 80-kDa csA, is involved in EDTA-resistant cell contact at the aggregation stage of Dictyostelium discoideum. A 31-kDa csA was isolated from the 80-kDa csA by treatment with Achromobacter protease I. Results from thin-layer chromatography and MALDI-TOF MS analysis indicated that the 31-kDa csA contains ceramide as a component of glycosylphosphatidyl-inositol (GPI). Comparison between the 80-kDa csA and the 31-kDa csA treated with phosphatidylinositol-specific phospholipase C (PI-PLC) or GPI-specific phospholipase D (GPI-PLD) was carried out. Our results indicated that the GPI-anchor of the 31-kDa csA was more sensitive to PI-PLC treatment than that of the 80-kDa csA, and that the anchor in both was easily cleaved by GPI-PLD treatment. They suggested that the resistance of 80-kDa csA to PI-PLC treatment was due to steric hindrance and myo-inositol modification. The results of the 80-kDa csA and the 31-kDa csA treated with sphingomyelinase were similar to those with PI-PLC treatment. In the presence of 1,10-phenanthroline, a GPI-PLD inhibitor, development of Dictyostelium was markedly inhibited, suggesting that GPI-PLD is functional in developmental regulation through cell adhesion.
Yoshida
Motonobu
M
Department of Agriculture, Kinki University, Nakamachi, Nara 631-8505, Japan. yoshida_m@nara.kindai.ac.jp
Sakuragi
Naoya
N
Kondo
Ken
K
Tanesaka
Eiji
E
D000818:Animals; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D004023:Dictyostelium / Q000737:chemistry / Q000254:growth & development; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D007294:Inositol / Q000378:metabolism; D010618:Phenanthrolines / Q000494:pharmacology; D043265:Phosphatidylinositol Diacylglycerol-Lyase / Q000378:metabolism; D054801:Phosphoinositide Phospholipase C; D010739:Phospholipase D / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology; D010740:Phospholipases / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*
D015815:Cell Adhesion Molecules; D017261:Glycosylphosphatidylinositols; D010618:Phenanthrolines; D015800:Protozoan Proteins; C052291:cell cohesion molecule, Dictyostelium; D007294:Inositol; D010740:Phospholipases; D054801:Phosphoinositide Phospholipase C; D010739:Phospholipase D; C065017:glycoprotein phospholipase D; D043265:Phosphatidylinositol Diacylglycerol-Lyase; C025205:1,10-phenanthroline