Molecular characteristics and peptide specificity of bradykinin binding sites in intact neuroblastoma-glioma cells in culture (NG 108-15).
Here we report that the mouse neuroblastoma-glioma hybrid cell line NG108-15 possess high-affinity binding sites for the nonapeptide bradykinin, as revealed by competitive displacement of 125I-8Tyr bradykinin by various bradykinin analogs. These binding sites were further characterized by covalent cross-linking of 125I-8Tyr bradykinin to intact NG108-15 grown as a monolayer, using dithiobis-succinimidylpropionate (DTSP) as a cross-linking reagent. Sodium dodecyl sulfate (SDS) electrophoresis after solubilization of the cross-linked cells, demonstrated the preferential and specific labeling of two polypeptides with apparent molecular weights of Mr = 36,000 and Mr = 47,000. A third polypeptide of Mr = 69,000 was labeled less intensely.
Lee
R T
RT
Laboratory of Cell Biology, National Institutes of Mental Health, Bethesda, Md 20892.
Lolait
S J
SJ
Muller
J M
JM
D000818:Animals; D001345:Autoradiography; D001667:Binding, Competitive; D001920:Bradykinin / Q000031:analogs & derivatives* / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000378:metabolism; D005910:Glioma / Q000378:metabolism*; D051379:Mice; D009447:Neuroblastoma / Q000378:metabolism*; D018002:Receptors, Bradykinin; D017981:Receptors, Neurotransmitter / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D003432:Cross-Linking Reagents; D018002:Receptors, Bradykinin; D017981:Receptors, Neurotransmitter; C026201:bradykinin, Tyr-; D001920:Bradykinin