Semi-Synthesis of Ion Channels

Creative Bioarray is committed to providing clients with semi-synthetic services of ion channels through protein trans-splicing, providing a powerful means for an in-depth understanding of the detailed functions and pharmacological characteristics of ion channels.

Background

Ion channels, transport proteins, and pumps constitute a large class of membrane proteins responsible for maintaining ionic homeostasis across cell membranes, which in turn play important roles in a variety of physiological processes. Traditional site-directed mutagenesis techniques and genetic code amplification methods have led to remarkable progress in understanding detailed functional and pharmacological insights of membrane proteins such as ion channels as well as developing therapeutics targeting these membrane proteins.

Recently, an increasing understanding of the function of ion channels has been complemented by semisynthetic strategies. Although the semisynthetic strategy is challenging, this strategy makes it possible to expand the type and number of modifications in ion channel research. Semisynthetic approaches have unique advantages over other approaches, of particular interest is protein trans-splicing (PTS), which has been seen as a powerful tool for protein modification or engineering.

Fig. 1 P2X2 receptor semi-synthesis. (Sarkar, 2021)

Our Services

Our research team provides clients with semi-synthetic ion channels and incorporates a wide range of chemical modifications to part of the protein that is generated synthetically, such as backbone modifications, chemical handles, non-canonical amino acids (ncAA), fluorescent or spectral labels, and any combination of these. Protein trans-splicing (PTS) and tandem protein trans-splicing (tPTS) methods are our common strategies.

• Constructs design for PTS/tPTS experiment.
  We provide our clients with the overall construct design by comprehensively considering the choice of split inteins, length and properties of peptide X, choice of splice sites, and generating and recombinantly testing the design of intein insertion sites.
• Design and synthesis of peptide X.
• Expression and optimization of recombinant N & C constructs with peptide X.

Through the above steps, we provide our clients with semi-synthetic services of various ion channels, such as the Na_v1.5 channel, KcsA, and P2X2 receptor. We have made important contributions to clients in the following projects:

• Insertion of recombinant N-terminal fragment in the acid-sensing ion channel (ASIC) 1a intracellular domain via PTS.
• Insertion of stable posttranslational modification (PTM) mimics into Na_v1.5 in Xenopus laevis oocytes via tPTS.
• Insertion of ncAAs in the P2X2 receptor extracellular domain via tPTS.

Our research team will improve the experimental strategy according to your scientific needs and formulate the best experimental scheme for you.

Advantages

• Complete instrument platform
• Exquisite experimental operation level
• Experience in customized solutions
• Competitive price

Creative Bioarray has rich experience in semi-synthetic ion channels. We are committed to providing customized solutions to meet the unique requirements of each client. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.

Reference

1. Sarkar, D.; et al. Ion channel engineering using protein trans-splicing//Methods in Enzymology. Academic Press, 2021, 654: 19-48.