Direct observation of individual KCNQ1 potassium channels reveals their distinctive diffusive behavior.

The Journal of Biological Chemistry
Gregory I MashanovAndrew Tinker

Abstract

We have directly observed the trafficking and fusion of ion channel containing vesicles and monitored the release of individual ion channels at the plasma membrane of live mammalian cells using total internal reflection fluorescence microscopy. Proteins were fused in-frame with green or red fluorescent proteins and expressed at low level in HL-1 and HEK293 cells. Dual color imaging revealed that vesicle trafficking involved motorized movement along microtubules followed by stalling, fusion, and subsequent release of individual ion channels at the plasma membrane. We found that KCNQ1-KCNE1 complexes were released in batches of about 5 molecules per vesicle. To elucidate the properties of ion channel complexes at the cell membrane we tracked the movement of individual molecules and compared the diffusive behavior of two types of potassium channel complex (KCNQ1-KCNE1 and Kir6.2-SUR2A) to that of a G-protein coupled receptor, the A1 adenosine receptor. Plots of mean squared displacement against time intervals showed that mobility depended on channel type, cell type, and temperature. Analysis of the mobility of wild type KCNQ1-KCNE1 complexes showed the existence of a significant immobile subpopulation and also a significant number...Continue Reading

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Citations

Nov 19, 2010·The Journal of Biological Chemistry·Alison M ThomasAndrew Tinker
Oct 22, 2010·The Journal of Biological Chemistry·Josef MadlGerhard J Schütz
Mar 5, 2014·Proceedings of the National Academy of Sciences of the United States of America·Leigh D PlantSteve A N Goldstein
Aug 23, 2011·Biochimica Et Biophysica Acta·Ikuomi MikuniWai-Meng Kwok
Aug 28, 2016·The Journal of Biological Chemistry·Thomas G BaboolalJustin E Molloy
Dec 17, 2020·The Journal of General Physiology·Gregory I MashanovJustin E Molloy

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