Characterization of Epithelial Sodium Channels

Creative Bioarray is committed to providing high-quality epithelial sodium channel (ENaC) characterization services to accelerate the understanding of the complex structure and important physiological functions of ENaCs. Our dedicated technical support and scientific services will contribute to your study of the physiopathological significance of these channels.

Introduction

ENaC is a member of the ENaC/DEG superfamily and is characterized by Na⁺ selectivity, voltage independence, and amiloride sensitivity. These channels work together to perform different cellular functions in different organisms. In humans, ENaCs are widely distributed in the apical membrane of epithelial tissues throughout the body, such as the respiratory tract, lungs, renal tubules, and placenta, and are involved in the regulation of systemic blood volume, salt, and water.

ENaCs typically assemble as a heterotrimer containing three subunits (α or δ, β, and γ) to form functional channels. However, after years of research, the number of subunits in active channels has not been clearly elucidated. It is necessary to further study the complex structure, biophysical properties, and functions of EnaCs using various methods such as patch clamp, immunolabeling, and quantitative PCR. These studies help to provide new ideas and methods for the subsequent exploration of therapeutic solutions, and provide the possibility of manipulating these channels to treat or prevent related diseases.

Fig. 1 Architecture of the human ENaC. (Noreng, 2018)

Our Services

We help our clients characterize the structure and function of ENaCs through multiple technologies. Our services include but not limited to:

• Subunit composition analysis

We help clients examine the subunit composition of ENaCs, determine whether subunits are cross-linked, and observe cross-linked complexes through a variety of biochemical techniques such as affinity chromatography and cation and anion exchange chromatography.

• Structure characterization of ENaCs

We help clients to determine the structure of ENaC in the uncleaved state and realize the cryo-EM three-dimensional reconstruction of ENaC through advanced biophysical technologies such as single particle cryo-electron microscope (cryo-EM).

• Determination of the positions of the GRIP domains.
• Analysis of domain-mediated unique interactions.
• Structural analysis of aromatic pocket formed by important gating domains in ENaC.

• Functional characterization of ENaCs
• Detection of plasma membrane localization of ENaCs using antibodies that bind to the extracellular domain of ENaCs.
• Detection of the selectivity of ENaC for Na⁺ and K⁺ and sensitivity for amiloride and trypsin treatment through whole-cell patch clamp electrophysiology and two-electrode voltage-clamp electrophysiology (TEVC).

Applications

• ENaC assembly research
• Study on the mechanism of ENaC gating
• Study on the pharmacological significance of ENaCs

After years of efforts, Creative Bioarray has accumulated rich experience in the functional and structural characterization of ion channels. We are confident that we can provide you with high-quality data related to ENaC structure and physiological function, opening new avenues for understanding the abnormal behavior of these channels that cause disease and developing treatment options. If you are interested in our services, please contact us for more details.

Reference

1. Noreng, S.; et al. Structure of the human epithelial sodium channel by cryo-electron microscopy. elife, 2018, 7: e39340.