rapid internalization of the oncogenic k+ channel kv10.1快速致癌k +通道kv10.1的内化.pdfVIP

Rapid Internalization of the Oncogenic K+ Channel KV 10.1 1¤ ¨ 1 1 ¨ 1,2 Tobias Kohl , Eva Lorinczi , Luis A. Pardo *, Walter Stuhmer * ¨ 1 Max-Planck-Institute of Experimental Medicine, Department of Molecular Biology of Neuronal Signals, Gottingen, Germany, 2 DFG Research Center for Molecular ¨ Physiology of the Brain (CMPB), Gottingen, Germany Abstract K 10.1 is a mammalian brain voltage-gated potassium channel whose ectopic expression outside of the brain has been V proven relevant for tumor biology. Promotion of cancer cell proliferation by KV 10.1 depends largely on ion flow, but some oncogenic properties remain in the absence of ion permeation. Additionally, KV 10.1 surface populations are small compared to large intracellular pools. Control of protein turnover within cells is key to both cellular plasticity and homeostasis, and therefore we set out to analyze how endocytic trafficking participates in controlling KV 10.1 intracellular distribution and life cycle. To follow plasma membrane KV 10.1 selectively, we generated a modified channel of displaying an extracellular affinity tag for surface labeling by a-bungarotoxin. This modification only minimally affected KV 10.1 electrophysiological properties. Using a combination of microscopy and biochemistry techniques, we show that KV 10.1 is constitutively internalized involving at least two distinct pathways of endocytosis and mainly sorted to lysosomes. This occurs at a relatively fast rate. Simultaneously, recycling seems to contribute to maintain basal K 10.1 surfa