Kv2.1 cell surface clusters are insertion platforms for ion channel delivery to the plasma membrane.
Voltage-gated K(+) (Kv) channels regulate membrane potential in many cell types. Although the channel surface density and location must be well controlled, little is known about Kv channel delivery and retrieval on the cell surface. The Kv2.1 channel localizes to micron-sized clusters in neurons and transfected human embryonic kidney (HEK) cells, where it is nonconducting. Because Kv2.1 is postulated to be involved in soluble N-ethylmaleimide-sensitive factor attachment protein receptor-mediated membrane fusion, we examined the hypothesis that these surface clusters are specialized platforms involved in membrane protein trafficking. Total internal reflection-based fluorescence recovery after photobleaching studies and quantum dot imaging of single Kv2.1 channels revealed that Kv2.1-containing vesicles deliver cargo at the Kv2.1 surface clusters in both transfected HEK cells and hippocampal neurons. More than 85% of cytoplasmic and recycling Kv2.1 channels was delivered to the cell surface at the cluster perimeter in both cell types. At least 85% of recycling Kv1.4, which, unlike Kv2.1, has a homogeneous surface distribution, is also delivered here. Actin depolymerization resulted in Kv2.1 exocytosis at cluster-free surface membrane. These results indicate that one nonconducting function of Kv2.1 is to form microdomains involved in membrane protein trafficking. This study is the first to identify stable cell surface platforms involved in ion channel trafficking.
Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Emily
E
Deutsch
Aubrey V
AV
Weigel
Elizabeth J
EJ
Akin
Phil
P
Fox
Gentry
G
Hansen
Christopher J
CJ
Haberkorn
Rob
R
Loftus
Diego
D
Krapf
Michael M
MM
Tamkun
D002462:Cell Membrane / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D015640:Ion Channel Gating; D051665:Kv1.4 Potassium Channel / Q000378:metabolism*; D008561:Membrane Fusion / Q000502:physiology; D008564:Membrane Potentials; D018613:Microscopy, Confocal; D009474:Neurons / Q000378:metabolism*; D050600:SNARE Proteins / Q000378:metabolism*; D051669:Shab Potassium Channels / Q000378:metabolism*; D013499:Surface Properties
D051665:Kv1.4 Potassium Channel; D050600:SNARE Proteins; D051669:Shab Potassium Channels