Reconstitution and partial purification of the sodium and chloride-coupled glycine transporter from rat spinal cord.
The (Na+ + Cl-)-coupled glycine transporter has been solubilized from rat spinal cord with 2% cholate and purified 6-7-fold using Wheat Germ Agglutinin-Sepharose 4B. Transport activity - as determined upon reconstitution of the fraction into liposomes - was retained on the column and eluted by N-acetylglucosamine. When the glycoprotein fraction was depleted of the N-acetylglucosamine and applied to a second round of lectin-chromatography, the glycine transport activity was retained and again could be eluted by the sugar. The transporter activity reconstituted from the glycoprotein fraction retains the same features displayed in the synaptic plasma membrane vesicles, namely an absolute dependence on sodium and chloride, electrogenicity and efflux and exchange properties. These observations indicate that the (Na+ + Cl-)-coupled glycine transporter is a glycoprotein.
Lopez-Corcuera
B
B
Departamento de Biología Molecular, Facultad de Ciencias, Universidad Autónoma, Madrid, Spain.
Kanner
B I
BI
Aragón
C
C
D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism*; D005998:Glycine / Q000378:metabolism*; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism; D008564:Membrane Potentials; D018377:Neurotransmitter Agents / Q000378:metabolism*; D011510:Proteolipids; D051381:Rats; D011919:Rats, Inbred Strains; D012964:Sodium / Q000378:metabolism*; D049992:Sodium Chloride Symporters; D012995:Solubility; D013116:Spinal Cord; D027981:Symporters; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D002352:Carrier Proteins; D002712:Chlorides; D006023:Glycoproteins; D018377:Neurotransmitter Agents; D011510:Proteolipids; D049992:Sodium Chloride Symporters; D027981:Symporters; D014909:Wheat Germ Agglutinins; C020485:proteoliposomes; D012964:Sodium; D005998:Glycine