Regulated recycling and plasma membrane recruitment of the high-affinity choline transporter.
The high-affinity choline transporter (CHT1) is responsible for uptake of choline from the synaptic cleft and supplying choline for acetylcholine synthesis. CHT1 internalization by clathrin-coated vesicles is proposed to represent a mechanism by which high-affinity choline uptake can be modulated. We show here that internalized CHT1 is rapidly recycled back to the cell surface in both human embryonic kidney cells (HEK 293 cells) and SH-SY5Y neuroblastoma cells. This rapidly recycling pool of CHT1 comprises about 10% of total CHT1 protein. In the SH-SY5Y neuroblastoma cell line K(+)-depolarization promotes Ca(2+)-dependent increase in the rate of CHT1 recycling to the plasma membrane without affecting the rate of CHT1 internalization. K(+)-depolarization also increases the size of the pool of CHT1 protein that can be mobilized to the plasma membrane. Thus, the activity-dependent increase in plasma membrane CHT1 localization appears to be regulated by two mechanisms: (i) an increase in the rate of externalization of the intracellular CHT1 pool; and (ii) the recruitment of additional intracellular transporters to the recycling pool.
Ribeiro
Fabiola M
FM
Cell Biology Research Group, Robarts Research Institute London, Ontario, Canada.
Pinthong
Metta
M
Black
Stefanie A G
SA
Gordon
Alexis C
AC
Prado
Vania F
VF
Prado
Marco A M
MA
Rylett
R Jane
RJ
Ferguson
Stephen S G
SS
D001667:Binding, Competitive; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D015151:Immunoblotting; D011189:Potassium Chloride / Q000494:pharmacology; D013194:Staining and Labeling; D027981:Symporters / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
C510731:SLC5A7 protein, human; D027981:Symporters; D011189:Potassium Chloride