Mechanisms through which gangliosides inhibit PDGF-stimulated mitogenesis in intact Swiss 3T3 cells: receptor tyrosine phosphorylation, intracellular calcium, and receptor binding.
Several potential mechanisms through which gangliosides could modulate PDGF-stimulated events in Swiss 3T3 cells were studied using intact cells. Of the gangliosides studied, at low micromolar concentrations GM2 was the only one that inhibited PDGF-stimulated DNA synthesis, but GT1b was the most potent between 25 and 100 microM; GM1 was generally the least effective, and GD1a and GM3 had intermediate effects. All gangliosides tested inhibited the PDGF-stimulated increases in free intracellular calcium concentrations ([Ca2+]i) with the rank order of potency being GM1 > or = GT1b > GM2 > GM3. PDGF stimulated phosphorylation on tyrosine of a protein with apparent M(r) = 170 kDa which was immunoprecipitated by an anti-PDGF receptor (beta) antibody, indicating that it is a PDGF receptor. Preincubating the cells with specific gangliosides inhibited tyrosine phosphorylation of this protein in a dose-responsive fashion with the following rank order of potency GD1a = GT1b > GM1 > GM2 > GM3. Autoradiography showed that this was due to a decrease in the proportion of cells synthesizing DNA, and a time study showed that ganglioside did not delay entry of the cells into S phase. These effects were not due to gangliosides interfering with PDGF binding to its receptor because results of competitive binding studies showed that none of the gangliosides studied had an effect on either receptor number or affinity, and did not bind to PDGF in solution. These results show that gangliosides affect several specific components of the complex responses to PDGF in intact cells. The relative effectiveness of individual gangliosides, however, varied among the different cellular and molecular responses. This is interpreted to mean that specific gangliosides modulate to different degrees several molecular mechanisms which converge on the common biological response of mitogenesis, and suggests that gangliosides as a family of molecules may function as coordinators of different molecular events involved in complex cellular processes.
Yates
A J
AJ
Division of Neuropathology, Ohio State University, Columbus 43210.
VanBrocklyn
J
J
Saqr
H E
HE
Guan
Z
Z
Stokes
B T
BT
O'Dorisio
M S
MS
D016475:3T3 Cells; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects*; D005732:Gangliosides / Q000494:pharmacology*; D051379:Mice; D010766:Phosphorylation; D010982:Platelet-Derived Growth Factor / Q000037:antagonists & inhibitors*; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D017479:Receptors, Platelet-Derived Growth Factor / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D005732:Gangliosides; D010982:Platelet-Derived Growth Factor; D014443:Tyrosine; D017479:Receptors, Platelet-Derived Growth Factor; D017027:Protein Tyrosine Phosphatases; D002118:Calcium