Histamine, actin-gelsolin binding, and polyphosphoinositides in human umbilical vein endothelial cells.
Histamine activates inositol phospholipid metabolism, increases calcium, and causes a change in shape of human umbilical vein endothelial (HUVE) cells. Changes in endothelial cell shape are determined, in part, by changes in the actin cytoskeleton. Gelsolin is an actin-binding protein with the potential to alter the actin cytoskeleton in response to changes in cell calcium and/or changes in polyphosphoinositides. Therefore, we examined the interactions of actin and gelsolin in HUVE cells in which inositol phospholipid metabolism was activated with histamine. In HUVE cells exposed to histamine we estimated actin-gelsolin binding by quantitating actin and gelsolin, immunoprecipitated with anti-gelsolin Sepharose. We estimated the relative amount of filamentous actin in the histamine-exposed HUVE cells by quantitating the amount of actin that was Triton soluble. We also measured the amount of phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) in the HUVE cells before and after exposure to histamine. We found that histamine decreased the amount of actin that was immunoprecipitated with gelsolin, decreased the fraction of cell actin that was Triton soluble, and increased PIP and PIP2. These results demonstrate that histamine promotes actin filament formation in HUVE cells and that histamine-mediated changes in actin-gelsolin binding in these cells are better predicted by changes in polyphosphoinositides than by increases in cell calcium.
Carson
M R
MR
Department of Medicine, University of Iowa College of Medicine, Iowa City 52242.
Shasby
S S
SS
Lind
S E
SE
Shasby
D M
DM
D000199:Actins / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D018260:Gelsolin; D006632:Histamine / Q000378:metabolism*; D006801:Humans; D007294:Inositol / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D011092:Polyethylene Glycols; D012995:Solubility; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism*
D000199:Actins; D002135:Calcium-Binding Proteins; D018260:Gelsolin; D008840:Microfilament Proteins; D010716:Phosphatidylinositols; D010743:Phospholipids; D011092:Polyethylene Glycols; D007294:Inositol; D006632:Histamine