Interaction Analysis of Ion Channels and Other Proteins

The functions of ion channels are exquisitely regulated by interacting proteins. Creative Bioarray is committed to providing clients with analytical services for ion channel macromolecular interactions to help them deduce multiple interactions within the same cell and quantify changes in binding due to cell signaling.

Background

Ion channels are macromolecular complexes whose functions are precisely regulated by a rich repertoire of regulatory proteins. Regulation of ion channels enables ion flux to be coordinated with subcellular precision to maintain multiple physiological roles in organisms. Disruption of this process is often closely associated with a variety of human diseases, including channelopathies. Therefore, it is necessary to deeply study ion channel assembly and dynamic interactions, which are crucial for understanding and elucidating the detailed physiological and pathophysiological mechanisms of ion channels.

In recent years, a variety of cell biology and biophysical methods have been applied to study ion channel assemblies and probe various facets of ion channel macromolecular complexes, including X-ray crystallography, single-particle cryo-electron microscopy, optical methods, super-resolution microscopy, and others. These methods have significant spatial and temporal resolutions. In comparison, fluorescence resonance energy transfer (FRET) exerts unique advantages in monitoring biological interactions in living cells, providing a powerful biological tool for analyzing the dynamic changes of ion channel interactions in living cells.

Fig. 1 Application of FRET 2-hybrid analysis to deduce stoichiometry and binding. (Rivas, 2021)

Our Services

• Analysis of the interaction between ion channels and proteins in vitro.
  We mainly use steady-state fluorescence spectroscopy to help our clients characterize protein-protein interactions in vitro. We focused on the development of various exogenous fluorescent probes such as dansyl calmodulin (D-CaM) to monitor the interaction of various proteins, including ion channels, with Ca²⁺-dependent regulatory protein calmodulin, as well as detect conformational changes as a consequence of interactions with Ca²⁺, peptides or proteins.
• Analysis of the interaction between ion channels and proteins in vivo.
  We help our clients track the dynamic changes of protein-protein interactions in vivo mainly through FRET measurements, which is a fast, low sample preparation requirement and can be obtained from various instrumentation. We have established the FRET methodology for deducing relative binding and stoichiometry of ion channel macromolecular complexes using flow cytometry. This method has the advantages of improving the throughput, reducing the sampling bias, and improving the accessibility of FRET experiments. We can help clients with a variety of analyses such as:
• Analysis of CaM interaction with the myosin Va and Ca_V1.2.
• Analysis of PKA regulation of CaVβ subunit binding to RGK proteins.

Advantages

• Unparalleled expertise and world-class technology platform
• Powerful and advanced data analysis tools
• Solutions tailored to your needs
• Cost-effective services

Creative Bioarray has rich experience in the interaction analysis of ion channels and other proteins. Our expertise provides a range of flexible solutions to meet your specific requirements and fit your budget. If you are interested in our services, please feel free to contact us.

Reference

1. Rivas, S.; et al. Probing ion channel macromolecular interactions using fluorescence resonance energy transfer//Methods in enzymology. Academic Press, 2021, 653: 319-347.