Regulation of thrombin receptors on human umbilical vein endothelial cells.
Activated thrombin receptors on human umbilical vein endothelial cells rapidly undergo homologous desensitization, leaving the cells unable to respond to thrombin. The present studies examine the fate of activated thrombin receptors on endothelial cells and the mechanisms that restore intact receptors to the cell surface. The results show that: 1) at biologically relevant concentrations, thrombin rapidly cleaves all of its receptors on the cell surface. 2) The cleaved receptors are cleared from the cell surface in a two-phase process, with 60% being internalized within 10 min, the remainder requiring several hours. 3) The restoration of intact, thrombin-responsive receptors on the cell surface initially occurs from an intracellular pool of receptors in a process that is independent of protein synthesis. 4) Recycling of cleaved receptors either does not occur on endothelial cells or is masked by receptor clearance. 5) Subconfluent endothelial cells re-express intact receptors on the cell surface at a slower rate than confluent cells. 6) The agonist peptide, SFLLRN, also causes receptor internalization, although at concentrations greater than those required for receptor activation and desensitization. These results are distinctly different from those observed with megakaryoblastic cell lines, where > 90% of the activated thrombin receptors are internalized rapidly, up to 40% of the cleaved receptors are recycled, and no intracellular pool of intact receptors has been detected. Since the primary structure of the thrombin receptor is the same in all the cell types studied, these results demonstrate that there can be substantial differences between cell types in the mechanisms involved in the clearance of activated receptors and the re-expression on the cell surface of intact receptors capable of responding to thrombin.
Woolkalis
M J
MJ
Department of Physiology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
DeMelfi
T M
TM
Blanchard
N
N
Hoxie
J A
JA
Brass
L F
LF
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D004705:Endocytosis; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology; D018179:Receptors, Thrombin / Q000819:agonists / Q000378:metabolism*; D014471:Umbilical Veins
D010446:Peptide Fragments; D018179:Receptors, Thrombin; C082835:thrombin receptor peptide (42-47)