Analysis of Inter-Domain Interactions In Ion Channels

Creative Bioarray is dedicated to providing clients with the analysis of inter-domain interactions in ion channels through various technologies, such as cysteine-based cross-linking methods. Our specialized scientific services are helpful to generate an important mechanistic understanding of the role of inter-domain interactions in coordinating ligand binding and mediating channel gating and ion permeation.

Background

With few exceptions, ion channels consist of multiple transmembrane subunits that are entwined with each other around a central aqueous ion-conducting pore. Examples include cysteine-loop receptors, ionotropic glutamate receptors, ATP-gated P2X receptors, and others. These functional domains generate complex ion channels through multiple interactions and allow control of channel behavior with high sensitivity and precision, with important implications for multiple properties of ion channels, including gating, permeability, pharmacology, and subunit modulation.

An increasing number of ion channel structures have been solved at the atomic level, and the acquisition of these high-resolution structures has provided useful information to reveal many interactions or contacts between domains located within the same or adjacent subunits. Structure-based prediction facilitates the development of strategies to probe the interaction between two adjacent domains or regions and determine their impact on the functional properties of ion channels. Therefore, understanding how domains interact in ion channels is critical to better understanding channel function.

Fig. 1 Detecting a hERG channel domain–domain interaction with FRET. (Zheng, 2013)

Our Services

Our research team has established a cysteine-based cross-linking method, which follows the same disulfide bond principle and involves the introduction of double cysteine substitution into two positions located on the opposite side of the interface between the two domains, or from adjacent subunits or within the same subunits. The cross-linking of substituted cysteines due to the formation of disulfide bonds changes the functional properties of this ion channel. Through this strategy, we have successfully helped our clients study the functional role of interaction or contact between two adjacent domains existing in the ion channel.

• Measurement of changes in the ion channel functions.
• Analysis of the interaction or contact between two adjacent domains.

In addition to the above strategy, we have also established a protocal to measure fluorescence resonance energy transfer (FRET) signals by combining fluorescence microscopy and spectrograph to analyze the inter-domain interactions of some ion channels, such as detecting the domain-domain interactions of a human ether-á-go-go–related gene (hERG) channel.

Advantages

• State-of-the-art facilities and experienced experimental technicians
• Fully developed and flexible strategies
• Fast delivery cycle of data results
• Years of experience in customized solutions

Creative Bioarray is dedicated to helping clients analyze inter-domain interactions in ion channels. Our innovative research team strives to continually develop and optimize strategies to meet your unique scientific needs. If you are interested in our services, please feel free to contact us.

Reference

1. Zheng, J. Domain–domain interactions in ion channels. Journal of General Physiology, 2013, 142(4): 347-350.