Analysis of Mechanically Activated Ion Channels

Creative Bioarray is committed to providing clients with analytical services for mechanically activated ion channels (MACs), including the characterization of the structure, function, and properties of these ion channels, providing an important basis for understanding the mechanical force sensing mechanisms.

Background

All organisms have the ability to sense physical forces. MACs are an important physiological basis for cells to perform mechanotransduction and convert mechanical stimuli into biochemical signals. The ability of organisms to respond and adapt to mechanical stimuli plays an important role in many physiological phenomena, such as the generation of hearing and balance, regulation of cell size and shape, and cell movement. In recent years, the analysis of new MAC families such as the PIEZO and OSCA/TMEM63 channels has facilitated further developments in the field of mechanotransduction.

Characterizing the structure and function of MACs is important for understanding their diversity and the underlying mechanisms of gating and ion permeation. However, since physical stimuli (force, membrane tension) are relatively difficult to measure and control, the analysis of MACs is more difficult than channels activated by other stimuli, such as voltage or ligands. Stretch (or pressure clamp) and poke (or cell indentation) are currently commonly used methods for mechanical stimulation. All validated MACs have robust responses to one or both assays. Therefore, the response to stretch and/or poke has become the standard for testing new MAC candidates.

Fig. 1 Structures of some mechanically activated ion channels. (Kefauver, 2020)

Our Services

We are committed to the development and establishment of new technologies to help clients analyze the structure and physiology of MACs. We have accumulated a lot of successful experience in the field of analysis of MACs and our services can be applied to a variety of MACs, including two-pore potassium channels (K2Ps), PIEZO1 and PIEZO2, OSCA/TMEM63 channels, transient receptor potential (TRP) channels, mechano-electrical transduction channel complex, and degenerin (DEG) and ASICs. Our services include, but are not limited to:

• Structural characterization of MACs.
  We help clients resolve the structures of different MACs in detergent and lipid nanodiscs through techniques such as single-particle cryo-electron microscopy (cryo-EM), providing information to identify multiple structural features that might confer mechanosensitivity to these channels.
• Functional characterization of MACs.
  We offer our clients a variety of strategies to characterize the function and properties of MACs, including channel number, ion selectivity, unitary conductance, stimulus threshold, and stimulus adaptation. Our strategies include:
• A combination of stretch and poke stimulation and patch-clamp electrophysiology.
• A combination of pillar arrays and whole-cell patch-clamp, suitable for analysis of MACs at the cell-substrate interface.
• Analysis of MAC gating kinetics (activation, deactivation, inactivation, recovery from inactivation).
• Identification of MAC new families.

Applications

• Development of increasingly refined models of mechanotransduction
• Study on the mechanical force sensing mechanisms
• Study on the role of mechanotransduction in physiology and disease

Equipped with advanced experimental equipment and an excellent research team, Creative Bioarray has made substantial progress in the identification and characterization of MACs. We are confident to provide clients with professional and comprehensive MAC analysis services. If you need our technical support, please feel free to contact us.

Reference

1. Kefauver, J. M.; et al. Discoveries in structure and physiology of mechanically activated ion channels. Nature, 2020, 587(7835): 567-576.