Specific ganglioside-cell protein interactions: a study performed with GM1 ganglioside derivative containing photoactivable azide and rat cerebellar granule cells in culture.
The incubation of cultured rat cerebellar granule cells with a photoreactive derivative of radiolabeled GM1 ganglioside, [3H]GM1(N3), followed by illumination, led to the specific association of ganglioside to cell proteins. After 30 min of incubation only a few out of the cell proteins became radiolabeled. Two of these, at apparent molecular weights of 95 and 112 kDa, are interacting with the portion of associated ganglioside that is released by trypsin treatment; others, in the region between 31 and 44 kDa, are probably bound to molecules of ganglioside inserted into the outer membrane layer, thus showing that the ganglioside association to the cell surface is a selective phenomenon, involving specific proteins. Increasing the incubation time up to 24 h resulted in a larger number of radiolabeled proteins, probably as a consequence of the internalization and metabolic processing of administered [3H]GM1(N3). In fact, photoreactive and radioactive metabolic derivatives of [3H]GM1(N3) can also interact with a number of proteins. After 24 h incubation, some radioactivity was also associated to cytosolic proteins. Again in this case the interaction with proteins seems to be a specific process involving only a few out of the total cytosolic proteins.
Sonnino
S
S
Department of Medical Chemistry and Biochemistry, Medical School, University of Milan, Italy.
Chigorno
V
V
Valsecchi
M
M
Pitto
M
M
Tettamanti
G
G
D000818:Animals; D001345:Autoradiography; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005677:G(M1) Ganglioside / Q000378:metabolism*; D005732:Gangliosides / Q000378:metabolism*; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D014316:Tritium
D005732:Gangliosides; D009419:Nerve Tissue Proteins; D014316:Tritium; D005677:G(M1) Ganglioside