title: Disposition of gangliosides and sialosylglycoproteins in neuronal membranes.
abstract: Labeled gangliosides and glycoproteins were obtained by incubation of homogenized neuronal perikarya from rat brain with CMP-[3H]N-acetyl neuraminic acid. The highest degree of labelling was observed in a subcellular fraction that also showed the highest specific activities for several ganglioside glycosyltransferases. The [3H]sialosylglycoconjugates of this fraction remained associated with the membranes after treatment with 1 M-KCl, 125 mM-EDTA, repeated freezing and thawing, or controlled sonication, but were solubilized by sodium deoxycholate (DOC) at a concentration high enough to solubilize the choline phospholipids. About 75% of th neuraminidase-labile sialosyl residues of these labeled endogenous gangliosides and glycoproteins were protected from the action of added neuraminidase or pronase or both enzymes added together. The protection was not abolished by pretreatment of the membranes with high ionic strength or with EDTA but was abolished by sonication or low concentration of DOC. Between 50 and 80% of the neuraminidase-labile sialosyl residues of the gangliosides of the neuronal perikaryon membrane fraction labelled in vivo by an intracerebral injection of N-[3H]acetylmannosamine were, at 3 h after the injection, also protected from the action of added neuraminidase. The protection was abolished by the addition of DOC. In contrast with behaviour of the labeled glycoconjugates of this neuronal perikaryon fraction, the gangliosides and sialosylglycoproteins from intact synaptosomes were accessible to neuraminidase. It is suggested that most gangliosides and sialosylglycoproteins are sialosylated as intrinsic components of the neuronal perikaryon membrane fraction and that at some stage of the process of transport through the axon and incorporation into the synaptic plasma membrane they change their accessibility to added enzymes.
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002540:Cerebral Cortex / Q000378:metabolism*; D005732:Gangliosides / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D009439:Neuraminidase; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D012795:Sialoglycoproteins / Q000378:metabolism*; D012799:Sialyltransferases / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D013574:Synaptosomes / Q000032:analysis; D014166:Transferases / Q000378:metabolism*
chemical_list: D005732:Gangliosides; D008563:Membrane Lipids; D008565:Membrane Proteins; D012795:Sialoglycoproteins; D014166:Transferases; D012799:Sialyltransferases; D009439:Neuraminidase