Dynamic regulation of β1 subunit trafficking controls vascular contractility.
Ion channels composed of pore-forming and auxiliary subunits control physiological functions in virtually all cell types. A conventional view is that channels assemble with their auxiliary subunits before anterograde plasma membrane trafficking of the protein complex. Whether the multisubunit composition of surface channels is fixed following protein synthesis or flexible and open to acute and, potentially, rapid modulation to control activity and cellular excitability is unclear. Arterial smooth muscle cells (myocytes) express large-conductance Ca(2+)-activated potassium (BK) channel α and auxiliary β1 subunits that are functionally significant modulators of arterial contractility. Here, we show that native BKα subunits are primarily (∼95%) plasma membrane-localized in human and rat arterial myocytes. In contrast, only a small fraction (∼10%) of total β1 subunits are located at the cell surface. Immunofluorescence resonance energy transfer microscopy demonstrated that intracellular β1 subunits are stored within Rab11A-postive recycling endosomes. Nitric oxide (NO), acting via cGMP-dependent protein kinase, and cAMP-dependent pathways stimulated rapid (≤1 min) anterograde trafficking of β1 subunit-containing recycling endosomes, which increased surface β1 almost threefold. These β1 subunits associated with surface-resident BKα proteins, elevating channel Ca(2+) sensitivity and activity. Our data also show that rapid β1 subunit anterograde trafficking is the primary mechanism by which NO activates myocyte BK channels and induces vasodilation. In summary, we show that rapid β1 subunit surface trafficking controls functional BK channel activity in arterial myocytes and vascular contractility. Conceivably, regulated auxiliary subunit trafficking may control ion channel activity in a wide variety of cell types.
Departments of Physiology and Neurosurgery, University of Tennessee Health Science Center, Memphis, TN 38163.
M Dennis
MD
Leo
John P
JP
Bannister
Damodaran
D
Narayanan
Anitha
A
Nair
Jordan E
JE
Grubbs
Kyle S
KS
Gabrick
Frederick A
FA
Boop
Jonathan H
JH
Jaggar
D000818:Animals; D001808:Blood Vessels / Q000502:physiology*; D031541:Fluorescence Resonance Energy Transfer; D017136:Ion Transport; D008297:Male; D018408:Patch-Clamp Techniques; D024681:Potassium Channels, Calcium-Activated / Q000737:chemistry / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D024681:Potassium Channels, Calcium-Activated
artery; protein trafficking