Clustering and Functional Coupling of Diverse Ion Channels and Signaling Proteins Revealed by Super-resolution STORM Microscopy in Neurons.
The fidelity of neuronal signaling requires organization of signaling molecules into macromolecular complexes, whose components are in intimate proximity. The intrinsic diffraction limit of light makes visualization of individual signaling complexes using visible light extremely difficult. However, using super-resolution stochastic optical reconstruction microscopy (STORM), we observed intimate association of individual molecules within signaling complexes containing ion channels (M-type K+, L-type Ca2+, or TRPV1 channels) and G protein-coupled receptors coupled by the scaffolding protein A-kinase-anchoring protein (AKAP)79/150. Some channels assembled as multi-channel supercomplexes. Surprisingly, we identified novel layers of interplay within macromolecular complexes containing diverse channel types at the single-complex level in sensory neurons, dependent on AKAP79/150. Electrophysiological studies revealed that such ion channels are functionally coupled as well. Our findings illustrate the novel role of AKAP79/150 as a molecular coupler of different channels that conveys crosstalk between channel activities within single microdomains in tuning the physiological response of neurons.
Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA.
Jie
J
Zhang
Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA.
Chase M
CM
Carver
Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA.
Frank S
FS
Choveau
Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA. Electronic address: shapirom@uthscsa.edu.
Mark S
MS
Shapiro
D054758:A Kinase Anchor Proteins / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D020746:Calcium Channels, L-Type / Q000378:metabolism* / Q000648:ultrastructure; D003412:Cricetulus; D005455:Fluorescent Antibody Technique; D006801:Humans; D051658:KCNQ2 Potassium Channel / Q000378:metabolism* / Q000648:ultrastructure; D051659:KCNQ3 Potassium Channel / Q000378:metabolism* / Q000648:ultrastructure; D008853:Microscopy; D046912:Multiprotein Complexes / Q000378:metabolism / Q000648:ultrastructure; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D061848:Optical Imaging; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism* / Q000648:ultrastructure; D050916:TRPV Cation Channels / Q000378:metabolism* / Q000648:ultrastructure
D054758:A Kinase Anchor Proteins; C487775:AKAP5 protein, human; D020746:Calcium Channels, L-Type; D051658:KCNQ2 Potassium Channel; D051659:KCNQ3 Potassium Channel; C419385:L-type calcium channel alpha(1C); D046912:Multiprotein Complexes; D043562:Receptors, G-Protein-Coupled; D050916:TRPV Cation Channels; C494482:TRPV1 protein, human
A-kinase anchoring proteins; ion channels; potassium channels; super-resolution microscopy
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