Characterization of Cyclic Nucleotide-Gated Ion Channels

Creative Bioarray is committed to providing clients with comprehensive and professional cyclic nucleotide-gated ion channels (CNG) characterization services. We not only help our clients obtain high-resolution structures about CNG channels, but also provide them with compositional and stoichiometric analysis as well as analysis of their biophysical and pharmacological properties.

Introduction

CNG channels are a class of channels composed of cyclic nucleotide-activated transmembrane proteins that play important roles in phototransduction and olfactory transduction, as well as neuronal pathfinding and synaptic plasticity. CNG channels are activated by the regulation of intracellular cyclic nucleotides cGMP or cAMP, and can non-selectively permeate cations such as Na⁺, K⁺, and Ca²⁺. These channels are expressed in a variety of tissues and cell types, including photoreceptors in the retina and olfactory receptor neurons.

Although CNG channel activity exhibits very little voltage dependence, CNG channels belong to the superfamily of voltage-gated ion channels. CNG channels typically form heterotetrameric complexes consisting of two or three different types of subunits. These channel complexes have distinct functional characteristics, such as ligand sensitivity and selectivity, ion permeability, and gating. Using a variety of techniques to study the functions of CNG channels, such as molecular structure, physiological and pathological roles, regulatory mechanisms, and gating mechanisms, will contribute to the prevention and treatment of CNG channel-related diseases.

Fig. 1 Proposed gating mechanism. (Mazzolini, 2018)

Our Services

Our research team has established a variety of experimental systems to help clients analyze the structure, function, and pharmacological properties of CNG channels. Our services include but not limited to:

• Analysis of the native assembly of CNG channels by functional and biochemical assays together with other methods, including fluorescence labeled subunits and fluorescence resonance energy transfer (FRET).
• Analysis of subunit stoichiometry and the spatial arrangement of subunits as heterotetramers by various strategies such as determining the effects of mutant subunits.
• Localization of subunits in neuronal cells and tissues as well as non-neuronal cells and tissues, such as the renal cortex, renal medulla, cardiac atrium, ventricle, testis, and spermatozoa.
• Structural characterization of CNG channels.
• Determination of the open and cGMP binding structure of CNG channels.
• Analysis of the conformational changes that occur in the cyclic nucleotide-binding domain (CNBD), C-linker, S6 region, and P region during channel gating.
• Monitoring of the gating movements of CNGs by the combination of various techniques, such as site-specific fluorescence and patch-clamp recording.
• Functional characterization of CNG channels by selective pharmacological tools.

Applications

• Development of specific pharmacological agents directed at CNG channels
• Study on the molecular mechanisms underlying CNG channel gating
• Development of new therapeutic strategies for CNG channel-related diseases

Creative Bioarray has extensive experience and expertise in ion channel analysis. Our dedicated research team will work with you to develop the best experimental protocol to deepen your understanding of the structure and function of CNG channels. If you are interested in our services, please contact us for more details.

Reference

1. Mazzolini, M.; et al. The gating mechanism in cyclic nucleotide-gated ion channels. Scientific reports, 2018, 8(1): 1-15.