title: Use of label-free optical biosensors to detect modulation of potassium channels by G-protein coupled receptors.
abstract: Ion channels control the electrical properties of neurons and other excitable cell types by selectively allowing ions to flow through the plasma membrane(1). To regulate neuronal excitability, the biophysical properties of ion channels are modified by signaling proteins and molecules, which often bind to the channels themselves to form a heteromeric channel complex(2,3). Traditional assays examining the interaction between channels and regulatory proteins require exogenous labels that can potentially alter the protein's behavior and decrease the physiological relevance of the target, while providing little information on the time course of interactions in living cells. Optical biosensors, such as the X-BODY Biosciences BIND Scanner system, use a novel label-free technology, resonance wavelength grating (RWG) optical biosensors, to detect changes in resonant reflected light near the biosensor. This assay allows the detection of the relative change in mass within the bottom portion of living cells adherent to the biosensor surface resulting from ligand induced changes in cell adhesion and spreading, toxicity, proliferation, and changes in protein-protein interactions near the plasma membrane. RWG optical biosensors have been used to detect changes in mass near the plasma membrane of cells following activation of G protein-coupled receptors (GPCRs), receptor tyrosine kinases, and other cell surface receptors. Ligand-induced changes in ion channel-protein interactions can also be studied using this assay. In this paper, we will describe the experimental procedure used to detect the modulation of Slack-B sodium-activated potassium (KNa) channels by GPCRs.
authors.affiliation: Department of Pharmacology, Yale School of Medicine.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Fleming
authors.affiliation: X-BODY Biosciences.
authors.forename: Steven M
authors.initials: SM
authors.lastname: Shamah
authors.affiliation: Department of Pharmacology, Yale School of Medicine; Department of Cellular and Molecular Physiology, Yale School of Medicine; leonard.kaczmarek@yale.edu.
authors.forename: Leonard K
authors.initials: LK
authors.lastname: Kaczmarek
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D055095:Optics and Photonics / Q000379:methods; D015221:Potassium Channels / Q000032:analysis / Q000378:metabolism*; D000081033:Potassium Channels, Sodium-Activated; D043562:Receptors, G-Protein-Coupled / Q000032:analysis / Q000378:metabolism*
chemical_list: C585122:KCNT1 protein, human; D009419:Nerve Tissue Proteins; D015221:Potassium Channels; D000081033:Potassium Channels, Sodium-Activated; D043562:Receptors, G-Protein-Coupled