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Originally published as MBC in Press, 10.1091/mbc.E02-06-0367 on December 7, 2002
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Vol. 14, Issue 3, 1149-1157, March 2003

Analysis of Na+,K+-ATPase Motion and Incorporation into the Plasma Membrane in Response to G Protein-coupled Receptor Signals in Living Cells

Alejandro M. Bertorello,*dagger Dagger Yulia Komarova,§dagger Kristen Smith,§@ Ingo B. Leibiger, Riad Efendiev,|| Carlos H. Pedemonte,|| Gary Borisy,§ and Jacob I. Sznajder#

 *Department of Medicine, Atherosclerosis Research Unit, and  Department of Molecular Medicine, Karolinska Institutet, Karolinska Hospital, S-171 76 Stockholm, Sweden;  §Department of Cell and Molecular Biology, and  #Division of Pulmonary and Critical Care Medicine, Northwestern University Medical School, Chicago, Illinois 60611;  ||College of Pharmacy, University of Houston, Houston, Texas 77204; and  @Laboratory of Cell Motility, A.N. Belozersky Institute, Moscow State University, Moscow, Russia

Dopamine (DA) increases Na+,K+-ATPase activity in lung alveolar epithelial cells. This effect is associated with an increase in Na+,K+-ATPase molecules within the plasma membrane (Ridge et al., 2002). Analysis of Na+,K+-ATPase motion was performed in real-time in alveolar cells stably expressing Na+,K+-ATPase molecules carrying a fluorescent tag (green fluorescent protein) in the alpha -subunit. The data demonstrate a distinct (random walk) pattern of basal movement of Na+,K+-ATPase-containing vesicles in nontreated cells. DA increased the directional movement (by 3.5 fold) of the vesicles and an increase in their velocity (by 25%) that consequently promoted the incorporation of vesicles into the plasma membrane. The movement of Na+,K+-ATPase-containing vesicles and incorporation into the plasma membrane were microtubule dependent, and disruption of this network perturbed vesicle motion toward the plasma membrane and prevented the increase in the Na+,K+-ATPase activity induced by DA. Thus, recruitment of new Na+,K+-ATPase molecules into the plasma membrane appears to be a major mechanism by which dopamine increases total cell Na+,K+-ATPase activity.


Dagger Corresponding author. E-mail address: aleber{at}mbox.ki.se.

Online version of this article contains video material. Online version is available at www.molbiolcell.org.

dagger Both authors contributed equally to this work.


Molecular Biology of the Cell
Vol. 14, 1149-1157, March 2003
Copyright © 2003 by The American Society for Cell Biology



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