Dopamine transporter: deglycosylation with exo- and endoglycosidases.
The dopamine transporter from rat caudate-putamen was photolabeled with [125I]DEEP as previously described. Treatment of photolabeled membranes with neuraminidase and N-glycanase reduced the molecular weight of the [125I]DEEP photolabeled dopamine transporter complex, whereas treatment with alpha-mannosidase had no effect. The solubilized [125I]DEEP photolabeled dopamine transporter complex readily bound to wheat-germ agglutinin but not to concanavalin-A sepharose columns. These results suggest that the carbohydrate moiety of the dopamine transporter is N-linked and contains significant quantities of sialic acid but not high mannose residues. A DEEP binding protein was readily detectable in other brain regions including the nucleus accumbens and olfactory tubercle, but not in the prefrontal cortex, olfactory bulb or hypothalamus under similar conditions. The DEEP binding protein in the other brain regions was similar to that in the striatum.
Lew
R
R
Neuroscience Branch, NIDA Addiction Research Center, Baltimore, MD 21224.
Grigoriadis
D
D
Wilson
A
A
Boja
J W
JW
Simantov
R
R
Kuhar
M J
MJ
D000818:Animals; D001386:Azides; D002241:Carbohydrates / Q000737:chemistry; D002352:Carrier Proteins / Q000737:chemistry*; D002846:Chromatography, Affinity; D003342:Corpus Striatum / Q000201:enzymology; D050483:Dopamine Plasma Membrane Transport Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases; D007457:Iodine Radioisotopes; D037102:Lectins; D008297:Male; D008361:Mannosidases; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D008970:Molecular Weight; D009419:Nerve Tissue Proteins; D009439:Neuraminidase; D009714:Nucleus Accumbens / Q000201:enzymology; D010777:Photochemistry; D010879:Piperazines; D051381:Rats; D011919:Rats, Inbred Strains; D043323:alpha-Mannosidase
D001386:Azides; D002241:Carbohydrates; D002352:Carrier Proteins; D050483:Dopamine Plasma Membrane Transport Proteins; D007457:Iodine Radioisotopes; D037102:Lectins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D010879:Piperazines; C060753:1-(2-(diphenylmethoxy)ethyl)-4-(2-(4-azido-3-iodophenyl)ethyl)piperazine; D006026:Glycoside Hydrolases; D008361:Mannosidases; C020668:glycanase; D009439:Neuraminidase; D043323:alpha-Mannosidase